Advantages at a glance
- Excellent SNP detection for reliable discrimination of different alleles
- Excellent performance, particularly suitable for allele-specific PCR (as-PCR) and also the first choice for highly selective PCRs
- Guaranteed simple primer design with standard primers where you only need to change the 3' nucleotide
- Guaranteed highly sensitive: less than 10 copies of a mutation can be detected in >10 000 wild-type copies
- No false amplification thanks to the aptamer-based hot-start function
- Free shipping
- Shipping within 24h
- 30 days money back guarantee
- Purchase comfortable on account
Product information "SNP Pol DNA Polymerase 2X PCR Mastermix"
The SNP Pol DNA polymerase > used for the SNP Pol 2X Mastermix has been specially designed for easy, reliable and rapid allele-specific discrimination, eg. CRISPR / Cas9 point mutations, for detecting incorrect CRISPR / Cas9 products, or validating sequencing results. The SNP Pol DNA polymerase distinguishes highly specific, whether a mismatch of the primer-template-complex is present or not. The mismatch (point mutation) must be at the 3 'end of the primer. Thus, mutant alleles can be distinguished exactly from wild-type alleles - without sequencing, since the polymerase simply does not amplify in the case of a mismatch.
Just place your primer on the supposed point mutation (Important: The point mutation must be at the 3 'end) and the polymerase will detect a mismatch in this region with almost 100% accuracy: If the template base complementary to the 3' end of the primer shows the mutation and the primer does not, no amplification takes place - 100% certainty within a short time!
The SNP Pol DNA polymerase can therefore be easily, time and cost-effectively used for the screening of point mutations.
The variant SNP PolTaq DNA polymerase has 5'-3 'nuclease activity and can therefore be used for specific hydrolysi probes such as Taqman® probes or Molecular beacons.
For more information on our SNP DNA polymerases and applications, read our blog now.
Description
SNP Pol DNA Polymerase and SNP PolTaq Polymerase are highly selective DNA polymerase for the detection of Single Nucleotide Polymorphisms. It was developed specifically for allele-specific discrimination, where a very high discrimination rate is required e.g., in allele-specific PCR (ASA; AS-PCR), allele-specific primer extension (AS-PEX), SNP analysis, genotyping or in methylation-specific PCRs (MSP). Many other DNA polymerases tolerate mismatched primer-template complexes and are therefor not suitable. The SNP Pol DNA polymerase, on the other hand, distinguishes these specifically (high discrimination) and supplies only PCR products with perfectly matching primer pairs! The Genaxxon SNP Pol and SNP PolTaq DNA polymerases differ up to 100% by means of allele-specific PCR between the two alleles and, after a simple qPCR, give a clear result as to which allele is present. Thus, the principly great potential of the CRISPR/Cas9 technology can be used for human medicine and plant biotechnology especially together with the SNP PolTaq or SNP Pol DNA polymerase.
Allele-specific PCR can be used to quantify the mutation rate in a pool or background of wild-type sequences. The verification of mutation frequencies determined by NGS can also be verified by means of allele-specific PCR and SNP Pol DNA polymerase. The SNP PolTaq DNA polymerase is very suitable for the analysis of liquid biopsy samples. With SNP PolTaq, the presence and frequency of cancer mutations can be analyzed and quantified very well.
Picture below: Application note SNP Pol DNA Polymerase
We recommend short amplicon length (about 60-200 bp) for best results, but also longer amplicon lengths are possible. The addition of additional Magnesium (+0.5 - 1.5mM) might be needed in case of longer amplicons >500 bp.
Trouble shooting:
No bands after PCR of 30 cycles!
Optimisation procedure for missing or weak bands.
Annealing temperature is too low!
Attention: The SNP Pol is an aptamer inhibited DNA polymerase. The aptamer oligo used reversibly inhibits the polymerase at temperatures <55°C! Therefore, the primers should ideally have an annealing temperature of >57°C.
- realtime PCR approach
- Check the dNTP concentration. This should be between 200 and 300µM (in the PCR).
- Increase the number of cycles (at least 35, preferably equal to 40)
Test optimisation - number of cycles
In endpoint detection, a cycle count of 30 is not always sufficient to generate a clearly visible band. For example, with less than 200 DNA copies as a starting value. The test should therefore be run using real-time PCR, or five parallel PCRs should be used, one of which is taken from the PCR device after each of five different cycles (example below).
Endpoint detection:
Set up five identical PCR reactions in parallel using the SNP Pol DNA polymerase.
Remove one of the PCR tubes from the cycler at each of 20, 25, 30, 35 and 40 cycles and apply all five reactions to an agarose gel at the end.
This makes it easy to recognise which is the optimum number of cycles in the PCR for the given application (starting material, target, primer).
SNP Pol PCR with cell lysates
Animal cells and E.coli can be used directly for PCR with the SNP Pol DNA polymerase! No separate lysis or proteinase K digestion is necessary.
PCR procedure:
1. 50 - 500 cells are required per PCR batch!
2. prepare PCR mix with primers and buffer and place on ice!
3. add the picked colony directly to 10-20µL water (no separate lysis and no proteinase K digestion),
vortex briefly (5 seconds), then add this cell suspension directly to the PCR reaction mixture, e.g.
10µL cell suspension for a PCR preparation with a total volume of 20µL. An initial denaturation time of 2-3 minutes is sufficient
is sufficient, can be extended to 5 minutes if necessary.
Genomic DNA per reaction max. 100 copies is relatively low, but should still work.
The remainder of this cell suspension can also be frozen away in order to carry out further tests.
Optional:
4. if possible: do real-time PCR!
The SNP Pol DNA polymerase (M3009 > or M3061 >) can be used together with non-specific fluorescent dyes (eg Genaxxon's Green DNA Dye > or SybrGreen®) in real-time PCR. When working with specific PCR probes, only the SNP PolTaq DNA polymerase can be used, since only these have a 5'-3 'exonuclease activity.
With our high quality dNTPs as Set (M3015.4100) > or Mix (M3016.1010) > or our DNALadders > and our favourable standard agarose (M3044) > we can offer additional products for your PCR.
Application areas for SNP Pol DNA and SNP Pol DNA polymerase
- Monitoring, verification and detection of point mutations
- Identification of correct or wrong CRISPR/Cas9 products
- Verification/validation of sequencing results
- Quantification of mutations (e.g. NGS results)
- SNP-detection by allele-specific amplification (ASA) / Allele-specific PCR
- Methylation specific PCRs (MSP) after bisulfite treated DNA (CpG methylation sides)
- HLA genotyping
- micro sequencing
- realtime PCR with hydrolysis probes
- realtime multiplex PCRs
- DamID-seq data in C. elegans.
As an alternative to ChIP, DNA adenine methyltransferase identification by sequencing (DamID-seq) was recently shown to be able to characterize binding sites in single mammalian cells. Additionally, DamID can be achieved for cell-type specific analysis by expressing Dam fusion proteins under tissue specific promoters in a controlled manner. In this report, we present a user-friendly pipeline to analyse DamID-seq data in C. elegans.
Sharma R, Ritler D, Meister P.
Tools for DNA adenine methyltransferase identification analysis of nuclear organization during C. elegans development. Genesis. 2016 Feb 4. doi: 10.1002/dvg.22925.
- Minisequencing SNP genotyping with SNPase DNA Polymerase can be carried out by the procedure described in:
Lovmar L, Fredriksson M, Liljedahl U, Sigurdsson S, Syvänen AC.
Quantitative evaluation by minisequencing and microarrays reveals accurate multiplexed SNP genotyping of whole genome amplified DNA. Nucleic Acids Res. 2003;31:e129.
- Allel specific mismatch selectivity by the HiDi DNA polymerase
Specifications:
2-time master mix. Contains all components necessary for direct PCR or PCR based genotyping.
SNP Pol DNA polymerase shows no 5’-3’ exonuclease activity! Not applicable for usage together with hydrolysis probes like, e.g. TagMan™ probes.
Attention: The SNP Pol is an aptamer inhibited DNA polymerase. The aptamer oligo used reversibly inhibits the polymerase at temperatures <55°C! Therefore, the primers should ideally have an annealing temperature of >57°C.
Application:
- CRISPR/Cas9 controls - validation of NGS sequencing results - SNP-detection by allele-specific amplification (ASA) / Allele-specific PCR - allelspezfische primer extension (AS-PEX) - Methylation specific PCR (MSP) - HLA genotyping - Micro-sequencingSource:
rec. from E.coliClassification:
Documents:
Safety Data SheetProtocols
Certificate
Manuals
Spec. Product Description
General Data 1
General Data 3
Category List
Source: NCBI PubMed
Matheus M. Noguera, Aimone Porri, Isabel S. Werle, James Heiser, Frank Brändle, Jens Lerchl, Brent Murphy, Michael Betz, Fanny Gatzmann, Martin Penkert, Clara Tuerk, Lucie Meyer, Nilda Roma-Burgos
Planta. 2022 Aug; 256(3):57.
doi: 10.1007/s00425-022-03968-2.
PMCID: PMC9374794.
Optimizing SNP Analysis with Aptamer-Based Inhibited DNA Polymerases: Key Tips for Successful PCR
Highly selective SNP DNA polymerases: The gold standard for allele-specific PCR testing and genotyping
SNP genotyping and its applications
Überprüfung und Kontrolle von Indels mit SNP Pol DNA-Polymerase
For better taste and flavor - Use SNP PolTaq for the unambiguous identification of point mutations, such as in Sake!
Accessory Items - Zubehör
White 384-Well-PCR-plates from whith polypropylene for optimum compatibility with the Roche LightCycler® 480 with 384 well block. The wells are designed for a working volume of up to 30µL.Properties:Virgin white polypropylenecut corner A24/P24up to 30µL working volume at 55µL total well capacity Key Features:- Molded from virgin polypropylene under ISO certified cleanroom conditions.- Free from DNase, RNase, human genomic DNA, and endotoxin/pyrogen.- Ultra-smooth, uniform, thin-walled wells for optimum PCR or RT-qPCR results.- Working volume: <30µL and a total well capacity of 55µL.- Maximum thermal conductivity for efficient heat transfer and precise thermal cycling.- Raised well rims prevent cross contamination and facilitate effective sealing for reduced evaporation.- Recommended for low volume PCR.- Compatible with blocks designed for 384 well PCR plates.- Compatible with standard multichannel pipettes.
Standard half skirted, low profile, 96-well PCR plate for Roche LC480. Thin walled for optimal heat transfer producing maximal PCR results. Sealing by film or caps, Fully certified for PCR use. These plates can also be used in an ABi 9700 cycler. Properties of plate material: Virgin Polypropylene, Surface characteristics optimised to reduced sample lose. Flexibility for variety of sealing methods. Continued performance after PCR at storage temperatures. Batch to batch consistency to maintain the same performance. Low profile improves low volume PCR. Black grid printed for easy identification. Cut off corner for fast orientation. White polypropylene.Volume: 100µL
The better performance of the Genaxxon bioscience 5X B-Enhancer Solution compared to standard enhancers such as form amide, DMSO, TMCA or BSA especially when used with GC rich regions or templates with a high degree of secondary structures depends on its ability to lower the DNA melting temperature and its enhancing effect on the DNA polymerase. In detail, Betaine binds preferentially to AT rich sequences in the major groove, thereby stabilizing AT rich regions of the DNA. Because AT forms 2 hydrogen bonds and GC forms 3, the bonding of AT is less stable than the one of GC. As a consequence of the stabilizing effect of Betaine on AT bonding, the stability of AT bonding and GC bonding is brought close to an equal level. At the same time, Betaine has a sequence independent destabilizing effect on all DNA. Summarized, the Tm of AT rich and GC rich sequences are equalized and the overall Tm is lowered. Furthermore, Betaine aids the processivity of thermostable polymerases and reduces "pauses" in polymerization caused by secondary structure that can induce the polymerase to disassociate from the DNA strand.As Betaine has a decreasing effect on the melting temperature of DNA and primers the denaturation temperatures as well as primer annealing temperatures should be reduced by 1 – 5°C compared to the annealing temperature already used. The optimal annealing temperature should be determined individually for each reaction.
Biotin-11-dUTP (Biotin-11-2'-deoxyuridin-5'-triphosphat - Tetralithiumsalt) is a commonly used component for non-radioactive labelling of DNA. Biotin-11-dUTP can be incorporated into DNA enzymatically by Nick-Translation, random-priming, 3'-end terminal labelling or by PCR. The figure '11' stands for the number of carbon atoms of the linker between dUTP and Biotin. The longer the 'linker-chain' the better the interaction between Biotin and Avidin. The shorter the linker the better the incorporation of dUTP into DNA. '11' has been shown to be a good compromise for most applications. The product Biotin-11-dUTP for non-radioactive labelling of DNA is delivered as 1 mM solution in 10 mM Tris-HCl (pH7.5), 1 mM EDTA. Biotin-11-dUTP can be enzymatically incorporated into DNA with Taq DNA Polymerase >, phi29 DNA Polymerase, Klenow Fragment, and DNA Polymerase I. Besides Biotin-11-dUTP Genaxxon offers other modified nucleotides or non modified dNTPs as our PCR dNTP-Mix> with 2 mM > or 10 mM > respective our dNTP-Set 100mM >. You can get additional products for your successful PCR as our Taq DNA Polymerse (M3001) >, or our proof-reading polymerases Pfu (M3004) >, Pwo (M3002) > and ReproFast (M3003) >, as well as the ready-to-use RedMastermix (M3029) >, already including dNTPs and a red dye.
This plate is compatible with 96 well block cycler designed for skirted plates. The plate is made from polypropylene with low profile 0.15mL wells, black grid reference, and a cut corner at H1. Benefits: Maximum thermal conductivity for efficient heat transfer and precise thermal cycling. Raised well rims prevent cross contamination and facilitate effective sealing for reduced evaporation. Super clear wells maximise sample visibility. Low profile plates with a working volume of 150µL and a total volume of 200µL when used with sealing options. The I2010 and I2261 plate fits, e.g. into the qTower 2.0 from Analytik Jena or the LabChip GXII Touch HT from Perkin Elmer.
Deoxyuridine 5'-triphosphate, tetrasodium salt (dUTP) of purity >99% (HPLC). dUTP may be used in place of dTTP in PCR and RT-PCR protocols to prevent carryover from previous amplifications. Product has been tested for PCR products of length up to 8kb (Taq DNA polymerase) and 0.8kb (Pfu DNA polymerase). The substitution of dTTP for dUTP in PCR results in uracil-containing PCR products that are suitable for most standard applications. The enzyme Uracil-DNA-Glycosylase, UDG >, can be added to a PCR premix to excise uracil from any contaminating PCR product, thereby preventing false positives. Genaxxon offers also PCR dNTP-Mix 10 mM >, unser dNTP-Set as 4 x 100 mM solution > or modified nucleotides as Biotin-11-dUTP >. For your successful PCR you will find different enzymes or PCR master mixes: Taq DNA Polymerse (M3001) >, Proof-Reading Polymerases Pfu (M3004) >, Pwo (M3002) > and ReproFast (M3003) >, as well as our ready-to-use RedMastermix (M3029) >, containing dNTPs already.
Green DNA Dye is a sensitive fluorescent dye for detection of dsDNA in qPCR (real-time PCR) and other applications. Because Green DNA Dye has greater sensitivity for dsDNA, it is especially useful for assays where the presence of contaminating RNA or ssDNA might obscure results. With exceptionally low background fluorescence Green DNA Dye is ideal for use with laser scanners. In the detection of double-stranded DNA in native polyacrylamide gels it is also much more sensitive than silver staining. When Green DNA Dye is free in solution, it emits a very low fluorescent signal. As soon as the dye binds to the double-stranded DNA, the signal increases significantly (thousand fold), which makes the fluorescent signal of the dye directly proportional to the amount of amplified dsDNA. Green DNA Dye is delivered as 100-time solution in DMSO. The concentration of Green DNA Dye is traditionally given as a dilution factor where the 1X dilution is used for staining of DNA gels. For PCR, a 0.2X dilution is the starting dilution for optimization. Our customers rely on Genaxxon's Green DNA Dye and use the dye for research into the regulation of tumor suppressor genes or vaccinations/disease patterns triggered by infections with enteroviruses, among other things. Preparation of a working solution for realtime PCR reactions:Green DNA Dye is suitable for real-time PCR with a suggested concentration for short targets of 0.2X (short targets: 50 – 400bp). Dilute delivered 100-time stock solution with nuclease-free PCR grade water by 1:10 (pre-dilution) and add 0.4µL of this solution to a 20µL final volume (PCR reaction). The 10-time working solution has to be kept at -20°C and has to be renewed every 2-3 weeks latest. Green DNA dye is a low toxic fluorescent nucleic acid dye as an alternative to ethidium bromide. Are you looking for more non-toxic dyes for your gel electrophoresis? Then try SafeGel red stain or SafeGel green stain - the cost-effective alternatives to EtBr. For qPCR we offer various qPCR Master Mixes > for your needs, with or without ROX, e.g., Green Master Mix High ROX >, Green Master Mix No ROX > or Probe Master Mix No ROX for qPCR >. Online form for your qPCR master mix test sample >
The Genaxxon qPCR adhesive seals are made of a optically clear adhesive film, with a pressure activated clue. The film is peelable and thus perfectly suitable for qPCR and other imaging techniques including crystallization applications. The adhesive is released after pressure on the surface. - Optically clear seal specifically developed for optical applications, particularly qPCR- It is non sticky when removed from the packaging; this aids handling when wearing gloves- The adhesive is contained within small capsules, allowing light to pass through to ensure the optical clarity of the seal- Seal integrity range: -40 °C to 110 °C- Free from DNase, RNase, and human genomic DNA When the seal is in position, pressure can be applied to burst the capsules, releasing a strong adhesive only where the seal touches the raised well rims of the plate - the rest of the seal area above the wells remains optically clear
ROX is used as passive reference dye to compensate for non-PCR related well to well variations in the fluorescence. Variations in fluorescence can occur e.g. due to well to well variation in light intensity, which depends on the optical construction of the PCR machine or due to pipetting variations. The ROX fluorescence does not change during the course of the PCR reaction but provide a stable baseline to which samples are normalized. The excitation and emission of the reference dye are 584nm and 612nm, respectively.The ROX concentration to be used depends on the Real Time PCR instrument or, more precisely, on the filter available. On earlier Real Time PCR machines there was no filter matching precisely to the ROX fluorescence. Therefore, high ROX concentrations are needed with these machines. Some newer machines do not need any internal standard to correct for light intensity. Nevertheless, an internal standard might still be useful to correct for pipetting variations.This dye has been qualified for use on instruments like the ABI PRISM® 7700, One-Step System, RotorGene and many others.Please consult the user manual of the real time PCR instrument with regard to appropriate levels of ROX. Ready to use qPCR Master Mixes with High ROX (500nM) >, Low ROX (50nM) > or without ROX > are also available from Genaxxon.
Clear, strongly adhesive sealing foils to peel off again. Applicable, e.g. for PCR plates / qPCR plates or other optical applications, e.g. fluorescence or colorimetric measurements. The film is not intended to be pierced with a pipette tip. - The sealing foils are made of a durable, transparent polyester film with a coating of strong adhesive.- The seals prevent cross over of individual samples and at the same time reliably prevention from evaporating.- The seals are suitable for normal PCR, and also for qPCR.- The seals can be used for PCR as well as for the storage of important samples. Removal: features adhesive-free end tabs for easy removal, will not leave a sticky residue on the plate surface following removal.
Uracil DNA Glycosylase (UDG), also called uracil N-glycosylase (UNG), catalyzes the release of uracil from uracil-containing DNA. The UDG/UNG from Genaxxon is a highly purified, recombinant enzyme isolated from E. coli with a mass of 26 kDa. UDG efficiently hydrolyzes single- and double-stranded DNA containing uracil, except for oligomers with 6 or fewer bases. Therefore, uracil-DNA glycosylase (UDG) is used wherever it is necessary to avoid the risk of DNA carryover (contamination) during PCR. In order to be able to hydrolyze DNA by UDG/UNG you do need dUTP > in your PCR reactions. Storage buffer: 20mM Tris HCl (pH 8.0) 50mM NaCl 0.1mM EDTA 50% Glycerin Reaction buffer (1X): 20 mM Tris HCl (pH 8.0) 1 mM EDTA
Accessory Items - related products
Lyo Ready qPCR MastermixA powerful, all-in-one solution for multiplex real-time PCR, optimized for high sensitivity, crude sample compatibility, and lyophilization. This highly concentrated 5x qPCR Master Mix is specifically designed for multiplex real-time PCR applications. It supports up to 30 targets in a single reaction, allows direct amplification from crude samples like blood or swabs (no extraction needed), and ensures high sensitivity with more space for primers and probes. Fully lyophilization-ready, it’s ideal for kit manufacturing, ambient shipping, and long-term storage without refrigeration.Key Benefits:Sensitive – More room for what mattersWith its 5x concentration, this Master Mix maximizes free volume for target-specific primers and probes. Ideal for high-level multiplexing without sacrificing sensitivity.Robust – Reliable results across targets5x qPCR Multiplex MasterMix ensures consistent, uniform amplification — even in complex multiplex panels.Fast Time to Result – No extraction neededWorks directly on crude samples like blood and swabs, eliminating the need for time-consuming DNA extraction steps.Specific – Precision built-inEngineered Taq DNA polymerase with enhanced room temperature stability and aptamer-based hot-start technology prevents non-specific amplification and enables a rapid start.Lyo Ready – Built for freeze-dryingFormulated with all necessary excipients for lyophilization. Can be freeze-dried in-house or by us. Once dried, it supports ambient storage and shipping — ideal for kit manufacturing and field use. Application notes:Learn more about how to use this product in practice – download the application note here.Read now in our blog why the 5x qPCR Multiplex MasterMix is the optimal choice for your multiplex qPCR. Our Standard Agarose LE and especially our high resolution Agarose Tiny are ideally suited for the subsequent electrophoresis analysis. Other realtime master mixes for your realtime PCR experiments can be found here .Examples of Multiplex applications: F. Javier Pérez-Pérez and Nancy D. Hanson, Detection of Plasmid-Mediated AmpC β-Lactamase Genes in Clinical Isolates by Using Multiplex PCR, J. Clin. Microbiol. June 2002 vol. 40 no. 6 2153-2162. doi: 10.1128/JCM.40.6.2153-2162.2002. Tamara B. Souzaa, Diego M. Lozerb, Sônia M. S. Kitagawab, Liliana C. Spanob, Neusa P. Silvac and Isabel C. A. Scaletskya, Real-Time Multiplex PCR Assay and Melting Curve Analysis for Identifying Diarrheagenic Escherichia coli. J. Clin. Microbiol. March 2013 vol. 51 no. 3 1031-1033, doi: 10.1128/JCM.02478-12.
Make a better choice! No choosing of different qPCR master mixes anymore! One product for use with different thermal cycler simplifies the decision making process. The GNX Versa qPCR Master Mix 2X is an optimized 2-time reaction mix for real-time qPCR detection and quantitation of target DNA sequences using the SYBR®/FAM channel of most real-time qPCR instruments. It contains our HotStart Taq DNA Polymerase together with a unique passive reference dye that is compatible across a variety of instrument platforms (including those that require a high or low ROX reference signal). This dye does not spectrally overlap with fluorescent dyes used for qPCR and will not interfere with real-time detection. The master mix formulation contains all PCR components required for amplification and quantitation of DNA except primers and DNA template. Genomic DNA or cDNA can be analysed using existing as well as commercial primers.AdvantagesOne formulation for different plattformsConvenient master mix formatUser-friendly protocols simplifing reaction setupProduct performs consistently across a wide variety of sample sourceData analysisSYBR® Green binds non-specifically to double-stranded DNA. For this, the best way to check if a qPCR reaction using SYBR® Green amplified a single target without primer dimer formation is with melt-curve analysis. Melt-curve analysis is performed after a qPCR run and serves to verify that PCR amplification was specific for a single amplicon. The nonspecific binding of SYBR® Green makes melt-curve analysis one of the very few ways to confirm target-specific amplification. If the fluorescence signal generated is due to primer dimers or non-specific amplification, your relative quantification of target is at least suspect. So when starting up a new SYBR® Green PCR, always perform melt-curve analysis as a QC step to help confirm target specificity!Consistent performance when using different instruments and modesStrong performance with multiple genes of various speciesRobustness against thawing and freezing cycles
GreenMastermix High ROX optimised for realtime PCR assays in block systems that contains all components to perform quantitative PCR with the exception of primer and template DNA. This 2X PCR master mix is ready-to-use and contains optimised amounts of all ingredients. Advantages of the chemically modified SuperHot Taq polymerase used in the Genaxxon GreenMasterMix: Hotstart technology enables setup of PCR mixture at room temperatur No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The chemical modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Green qPCR master mix high ROX contains all the necessary components in an optimized composition to carry out quantitative PCR: chemically modified Taq DNA Polymerase dATP, dCTP, dGTP, dTTP intercalating green fluorescent dye 500nM ROX optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) Stability at +2°C to +8°C (refrigerator): minimum of 8 months. This Mastermix is specially suited for the following instruments: Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900 HT, Eppendorf Realplex4, StepOne™ and StepOnePlus™. Other realtime PCR master mixes from Genaxxon are: M3023 - GreenMastermix No ROX >, M3045 - ProbeMastermix No ROX >, M3011 - GreenMastermix Low ROX >, M3031 - ProbeMastermix Low ROX >, M3052 - GreenMastermix High ROX >, M3010 - ProbeMastermix High ROX >.
GreenMastermix Low ROX optimised for realtime PCR assays in block systems that contains all components to perform quantitative PCR with the exception of primer and template DNA. This 2X Mastermix is ready-to-use and contains optimised amounts of all ingredients. Advantages of the chemically modified SuperHot Taq polymerase used in the Genaxxon GreenMasterMix: Hotstart technology enables setup of PCR mixture at room temperatur No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The chemical modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Green qPCR master mix high ROX contains all the necessary components in an optimized composition to carry out quantitative PCR: chemically modified Taq DNA Polymerase dATP, dCTP, dGTP, dTTP intercalating green fluorescent dye 50nM ROX optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) Stability at +2°C to +8°C (refrigerator): minimum of 8 months. This Mastermix is specially suited for the following instruments: Applied Biosystems® 7500, 7500 Fast and ViiA™ 7, QuantStudio™ instruments, Agilent Mx3000P™, Mx3005P™, Mx4000™ and AriaMx. Further realtime PCR Mastermixes and realtime FAST PCR Mastermixes can be found here: qPCR (Classic qPCR > or FAST and Multiplex qPCR >).
GreenMasterMix without ROX optimised for realtime PCR assays in block systems that contains all components to perform quantitative PCR with the exception of primer and template DNA. This 2X Mastermix is ready-to-use and contains optimised amounts of all ingredients. Advantages of the chemically modified SuperHot Taq polymerase used in the Genaxxon GreenMasterMix: Hotstart technology enables setup of PCR mixture at room temperatur No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The chemical modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Green qPCR master mix without ROX contains all the necessary components in an optimized composition to carry out quantitative PCR: chemically modified Taq DNA Polymerase dATP, dCTP, dGTP, dTTP intercalating green fluorescent dye optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) Stability at +2°C to +8°C (refrigerator): minimum of 8 months. This Mastermix is specially suited for the following instruments: BioRad CFX96 Touch™, CFX384 Touch™, CFX Connect™, DNA Engine Opticon® 2, Chromo4™, iCycler iQ™ and My iQ™ , Roche LightCycler® 480, LightCycler® 1536, LightCycler® Nano, LightCycler® 96 and QuantStudio™ instruments, Thermo Scientific™ PikoReal™, Cepheid SmartCycler®, Bio Molecular Systems Mic qPCR cycler, Qiagen Rotor Gene Q, Rotor Gene 6000, MyGo Mini and MyGo Pro. Further realtime PCR Mastermixes and realtime FAST PCR Mastermixes can be found here: qPCR (Classic qPCR > or FAST and Multiplex qPCR >).
GreenMasterMix FAST Blue with 500nM ROX is optimised for realtime PCR assays in block systems. This 2X Mastermix is ready-to-use and contains all components for a successful and reliable qPCR with the exception of primer and template DNA. Advantages of the HotStart Taq DNA polymerase used in the Genaxxon GreenMasterMix: Hotstart technology enables setup of PCR mixture at room temperature. short initial denaturing time of not more than 2 minutes No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The chemical modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Green qPCR master mix Blue with 500nM ROX contains all the necessary components in an optimized composition to carry out quantitative PCR: chemically modified Taq DNA Polymerase dATP, dCTP, dGTP, dTTP intercalating green fluorescent dye optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) This Mastermix is specially suited for the following instruments: Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900 HT, Eppendorf Realplex4, StepOne™ and StepOnePlus™. Other realtime PCR master mixes from Genaxxon are: M3023 - GreenMastermix No ROX >, M3045 - ProbeMastermix No ROX >, M3011 - GreenMastermix Low ROX >, M3031 - ProbeMastermix Low ROX >, M3052 - GreenMastermix High ROX >, M3010 - ProbeMastermix High ROX >.
GreenMasterMix FAST High ROX with 500nM ROX is optimised for realtime PCR assays in block systems. This 2X Mastermix is ready-to-use and contains all components for a successful and reliable qPCR with the exception of primer and template DNA. Advantages of the HotStart Taq DNA polymerase used in the Genaxxon GreenMasterMix: Hotstart technology enables setup of PCR mixture at room temperature. short initial denaturing time of not more than 2 minutes No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Green qPCR MasterMix High ROX contains all the necessary components in an optimized composition to carry out quantitative PCR: Hotstart Taq DNA Polymerase dATP, dCTP, dGTP, dTTP intercalating green fluorescent dye optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) This Mastermix is specially suited for the following instruments: Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900 HT, Eppendorf Realplex4, StepOne™ and StepOnePlus™. Further realtime PCR Mastermixes and realtime FAST PCR Mastermixes can be found here: qPCR (Classic qPCR > or FAST and Multiplex qPCR >).
GreenMasterMix FAST Low ROX with 50nM ROX is optimised for realtime PCR assays in block systems. This 2X Mastermix is ready-to-use and contains all components for a successful and reliable qPCR with the exception of primer and template DNA. Advantages of the HotStart Taq DNA polymerase used in the Genaxxon GreenMasterMix: Hotstart technology enables setup of PCR mixture at room temperature. short initial denaturing time of not more than 2 minutes No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Green qPCR master Low ROX contains all the necessary components in an optimized composition to carry out quantitative PCR: Hotstart Taq DNA Polymerase dATP, dCTP, dGTP, dTTP intercalating green fluorescent dye optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets 50nM internal passive reference fluorescence dye the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) This Mastermix is specially suited for the following instruments: Applied Biosystems® 7500, 7500 Fast and ViiA™ 7, QuantStudio™ instruments, Agilent Mx3000P™, Mx3005P™, Mx4000™ and AriaMx. Other realtime PCR master mixes from Genaxxon are: M3023 - GreenMastermix No ROX >, M3045 - ProbeMastermix No ROX >, M3011 - GreenMastermix Low ROX >, M3031 - ProbeMastermix Low ROX >, M3052 - GreenMastermix High ROX >, M3010 - ProbeMastermix High ROX >.
GreenMasterMix FAST without ROX is optimised for fast qPCR assays without probes in block systems. The master mix is optimized for fast PCR with short denaturation 2-step cycles. This 2X Mastermix is ready-to-use and contains all components for a successful and reliable qPCR with the exception of primer and template DNA. Advantages of the HotStart Taq DNA polymerase used in the Genaxxon GreenMasterMix: Hotstart technology enables setup of PCR mixture at room temperature. short initial denaturing time of not more than 2 minutes No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Green qPCR master mix without ROX contains all the necessary components in an optimized composition to carry out quantitative PCR: Hotstart Taq DNA Polymerase dATP, dCTP, dGTP, dTTP intercalating green fluorescent dye optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) Stability at +2°C to +8°C (refrigerator): minimum of 8 months. This Mastermix is specially suited for the following instruments: BioRad CFX96 Touch™, CFX384 Touch™, CFX Connect™, DNA Engine Opticon® 2, Chromo4™, iCycler iQ™ and My iQ™ , Roche LightCycler® 480, LightCycler® 1536, LightCycler® Nano, LightCycler® 96 and QuantStudio™ instruments, Thermo Scientific™ PikoReal™, Cepheid SmartCycler®, Bio Molecular Systems Mic qPCR cycler, Qiagen Rotor Gene Q, Rotor Gene 6000, MyGo Mini and MyGo Pro. Further realtime PCR Mastermixes and realtime FAST PCR Mastermixes can be found here: qPCR (Classic qPCR > or FAST and Multiplex qPCR >).
Multiplex PCR is a method that enables amplification of two or more amplicons simultaneously in a single reaction tube/reaction. It is widely used in genotyping and different areas of DNA testing in research, forensic and diagnostic laboratories. Our Multiplex HS Mastermix (2X) is an optimized ready-to-use mixture for probe-based assays such as TaqMan®, Beacons and MGBs. It contains a modified fast HotStart Taq DNA Polymerase, dNTPs and MgCl₂ combined in an optimized buffer system for realtime PCR / qPCR applications except primers, probe and template DNA / cDNA. The HotStart Taq Polymerase is based on the standard Taq DNA polymerase from Genaxxon inactivated by an specific antibody against Taq DNA polymerase which is activated by heat treatment. Thus, during setup and the first ramp of thermal cycling, the enzyme is not active and misprimed primers are not extended. The result is higher specificity, increased sensitivity and greater yields when compared to standard DNA polymerases, making this enzyme especially well-suited for multiplex PCR. - Amplification of multiple targets in a single tube- All-in-one master mix for convenient multiplexing- High specificity, sensitivity and product yield- Easy reaction setup at room temperature The Multiplex HS Master mix 2X is shipped in aliquots of 1mL. Our Standard Agarose LE > and especially our high resolution Agarose Tiny > are ideally suited for the subsequent electrophoresis analysis. Other realtime master mixes for your realtime PCR experiments can be found here >. Examples of Multiplex applications: F. Javier Pérez-Pérez and Nancy D. Hanson, Detection of Plasmid-Mediated AmpC β-Lactamase Genes in Clinical Isolates by Using Multiplex PCR, J. Clin. Microbiol. June 2002 vol. 40 no. 6 2153-2162. doi: 10.1128/JCM.40.6.2153-2162.2002. Tamara B. Souzaa, Diego M. Lozerb, Sônia M. S. Kitagawab, Liliana C. Spanob, Neusa P. Silvac and Isabel C. A. Scaletskya, Real-Time Multiplex PCR Assay and Melting Curve Analysis for Identifying Diarrheagenic Escherichia coli. J. Clin. Microbiol. March 2013 vol. 51 no. 3 1031-1033, doi: 10.1128/JCM.02478-12.
ProbeMastermix High ROX optimised for realtime PCR assays in block systems that contains all components to perform quantitative PCR with the exception of primer and template DNA. This 2X Mastermix is ready-to-use and contains optimised amounts of all ingredients. Multiplex PCR: Applications at Genaxxon and at customers site have shown that the qPCR Probe Mastermix can be used for the simultaneous detection of up to four DNA targets in the same PCR reaction. For further details please refer to the product manual or contact us: info@genaxxon.com. Advantages of the Genaxxon ProbeMasterMix: Hotstart technology enables setup of PCR mixture at room temperatur No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The chemical modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Probe qPCR master mix high ROX contains all the necessary components in an optimized composition to carry out quantitative PCR: chemically modified Taq DNA Polymerase dATP, dCTP, dGTP, dTTP 500nM ROX optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) Stability at +2°C to +8°C (refrigerator): minimum of 8 months. This Mastermix is specially suited for the following instruments: Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900 HT, Eppendorf Realplex4, StepOne™ and StepOnePlus™. Further realtime PCR Mastermixes and realtime FAST PCR Mastermixes can be found here: qPCR (Classic qPCR > or FAST and Multiplex qPCR >).
ProbeMasterMix low ROX optimised for realtime PCR assays in block systems that contains all components to perform quantitative PCR with the exception of primer and template DNA. This 2X master mix is ready-to-use and contains optimised amounts of all ingredients. Multiplex PCR: Applications at Genaxxon and at customers site have shown that the qPCR Probe Mastermix can be used for the simultaneous detection of up to four DNA targets in the same PCR reaction. For further details please refer to the product manual or contact us: info@genaxxon.com. Advantages of the Genaxxon ProbeMasterMix: Hotstart technology enables setup of PCR mixture at room temperatur No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The chemical modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Probe qPCR master mix without ROX contains all the necessary components in an optimized composition to carry out quantitative PCR. chemically modified Taq DNA Polymerase dATP, dCTP, dGTP, dTTP 50nM ROX optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) Stability at +2°C to +8°C (refrigerator): minimum of 8 months. This Mastermix is specially suited for the following instruments: Applied Biosystems® 7500, 7500 Fast and ViiA™ 7, QuantStudio™ instruments, Agilent Mx3000P™, Mx3005P™, Mx4000™ and AriaMx. Further realtime PCR Mastermixes and realtime FAST PCR Mastermixes can be found here: qPCR (Classic qPCR > or FAST and Multiplex qPCR >).
ProbeMasterMix FAST High ROX with 500nM ROX optimised for fast realtime PCR assays in block systems. This master mix is ready-to-use and contains all components for a successful and reliable quantitative PCR with the exception of primer and template DNA. Advantages of the Hotstart Taq polymerase used in the Genaxxon ProbeMasterMix: Hotstart technology enables setup of PCR mixture at room temperature. short initial denaturation of not more than 2 minutes. optimized for 2-step PCR protocols. No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon Hotstart Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The hotstart formulation inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Probe qPCR master mix High ROX with 500nM ROX contains all the necessary components in an optimized composition to carry out quantitative PCR. hotstart Taq DNA Polymerase dATP, dCTP, dGTP, dTTP optimized reaction buffer 500nM ROX as internal reference stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) This Mastermix is specially suited for the following instruments: Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900 HT, Eppendorf Realplex4, StepOne™ and StepOnePlus™. Further realtime PCR Mastermixes and realtime FAST PCR Mastermixes can be found here: qPCR (Classic qPCR > or FAST and Multiplex qPCR >).
ProbeMasterMix FAST Low ROX with 50nM ROX is optimised for fast qPCR assays with probes in block systems. The master mix is optimized for fast qPCR with short denaturation 2-step cycles. This 2X Mastermix is ready-to-use and contains all components for a successful and reliable qPCR with the exception of primer and template DNA Advantages of the Hotstart Taq polymerase used in the Genaxxon ProbeMasterMix: Hotstart technology enables setup of PCR mixture at room temperature. short initial denaturation of not more than 2 minutes. optimized for 2-step PCR protocols. No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon Hotstart Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The hotstart formulation inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Probe qPCR master mix with 50nM ROX contains all the necessary components in an optimized composition to carry out quantitative PCR. hotstart Taq DNA Polymerase dATP, dCTP, dGTP, dTTP optimized reaction buffer 50nM ROX as internal passive reference stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) This Mastermix is specially suited for the following instruments: Applied Biosystems® 7500, 7500 Fast and ViiA™ 7, QuantStudio™ instruments, Agilent Mx3000P™, Mx3005P™, Mx4000™ and AriaMx. Further realtime PCR Mastermixes and realtime FAST PCR Mastermixes can be found here: qPCR (Classic qPCR > or FAST and Multiplex qPCR >).
ProbeMasterMix FAST without ROX is optimised for fast qPCR assays with probes in block systems. The master mix is optimized for fast PCR with short denaturation 2-step cycles. This 2X Mastermix is ready-to-use and contains all components for a successful and reliable qPCR with the exception of primer and template DNA. Advantages of the Hotstart Taq polymerase used in the Genaxxon ProbeMasterMix: Hotstart technology enables setup of PCR mixture at room temperature. short initial denaturation of not more than 2 minutes. optimized for 2-step PCR protocols. No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon Hotstart Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The hotstart formulation inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Probe qPCR master mix without ROX contains all the necessary components in an optimized composition to carry out quantitative PCR. hotstart Taq DNA Polymerase dATP, dCTP, dGTP, dTTP optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1mL simplifies handling and storage (less freeze-thaw cycles per aliquote) This Mastermix is specially suited for the following instruments: BioRad CFX96 Touch™, CFX384 Touch™, CFX Connect™, DNA Engine Opticon® 2, Chromo4™, iCycler iQ™ and My iQ™ , Roche LightCycler® 480, LightCycler® 1536, LightCycler® Nano, LightCycler® 96 and QuantStudio™ instruments, Thermo Scientific™ PikoReal™, Cepheid SmartCycler®, Bio Molecular Systems Mic qPCR cycler, Qiagen Rotor Gene Q, Rotor Gene 6000, MyGo Mini and MyGo Pro. Further realtime PCR Mastermixes and realtime FAST PCR Mastermixes can be found here: qPCR (Classic qPCR > or FAST and Multiplex qPCR >).
ProbeMasterMix without ROX optimised for realtime PCR assays in block systems that contains all components to perform quantitative PCR with the exception of primer and template DNA. This 2X Mastermix is ready-to-use and contains optimised amounts of all ingredients. Multiplex PCR: Applications at Genaxxon and at customers site have shown that the qPCR Probe Mastermix can be used for the simultaneous detection of up to four DNA targets in the same PCR reaction. For further details please refer to the product manual or contact us: info@genaxxon.com. Advantages of the chemically modified SuperHot Taq polymerase used in the Genaxxon ProbeMasterMix: Hotstart technology enables setup of PCR mixture at room temperatur No pipetting on ice necessary anymore No immediate further processing (PCR) necessary. The pipetted pre-PCR mixtures can be left at RT for up to 3 days. Amplification of GC-rich templates High yields No Primer dimers The Genaxxon SuperHot Taq DNA polymerase offers convenience and practicality, without compromising efficiency or specificity: 1. The chemical modification inhibits the polymerase, which prevents false priming at all temperatures tested, allowing for PCR reactions to be set up at RT (20°C to 25°C), and 2. "High mix stability allows pre-assemby of the plate in advance", which means PCR reactions can be prepared and stored at RT for up to three days before they are run, without affecting the results. The Probe qPCR master mix without ROX contains all the necessary components in an optimized composition to carry out quantitative PCR. chemically modified Taq DNA Polymerase dATP, dCTP, dGTP, dTTP optimized reaction buffer stabilizers and enhancers to enable even amplification of low copy number targets the small aliquote size of 1.25mL simplifies handling and storage (less freeze-thaw cycles per aliquote) Stability at +2°C to +8°C (refrigerator): minimum of 8 months. This Mastermix is specially suited for the following instruments: BioRad CFX96 Touch™, CFX384 Touch™, CFX Connect™, DNA Engine Opticon® 2, Chromo4™, iCycler iQ™ and My iQ™ , Roche LightCycler® 480, LightCycler® 1536, LightCycler® Nano, LightCycler® 96 and QuantStudio™ instruments, Thermo Scientific™ PikoReal™, Cepheid SmartCycler®, Bio Molecular Systems Mic qPCR cycler, Qiagen Rotor Gene Q, Rotor Gene 6000, MyGo Mini and MyGo Pro. Further realtime PCR Mastermixes and realtime FAST PCR Mastermixes can be found here: qPCR (Classic qPCR > or FAST and Multiplex qPCR >).
SNP Pol DNA polymerase for easy, reliable and rapid allele-specific discrimination, eg. CRISPR / Cas9 point mutations, for detecting incorrect CRISPR / Cas9 products, or validating sequencing results. The SNP Pol DNA polymerase distinguishes highly specific, whether a mismatch of the primer-template-complex is present or not. The mismatch (point mutation) must be at the 3 'end of the primer. Thus, mutant alleles can be distinguished exactly from wild-type alleles - without sequencing, since the polymerase simply does not amplify in the case of a mismatch. Just place your primer on the supposed point mutation (Important: The point mutation must be at the 3 'end) and the polymerase will detect a mismatch in this region with almost 100% accuracy: If the template base complementary to the 3' end of the primer shows the mutation and the primer does not, no amplification takes place - 100% certainty within a short time!The SNP Pol DNA polymerase can therefore be easily, time and cost-effectively used for the screening of point mutations. The variant SNP PolTaq DNA polymerase > has 5'-3 'nuclease activity and can therefore be used for specific hydrolysi probes such as Taqman® probes or Molecular beacons. For more information on our SNP DNA polymerases and applications, read our blog now. DescriptionSNP Pol DNA Polymerase is a highly selective DNA polymerase for the detection of Single Nucleotide Polymorphisms. It was developed specifically for allele-specific discrimination, where a very high discrimination rate is required e.g., in allele-specific PCR (ASA; AS-PCR), allele-specific primer extension (AS-PEX), SNP analysis, genotyping or in methylation-specific PCRs (MSP). Many other DNA polymerases tolerate mismatched primer-template complexes and are therefor not suitable. The SNP Pol DNA polymerase, on the other hand, distinguishes these specifically (high discrimination) and supplies only PCR products with perfectly matching primer pairs! The Genaxxon SNP Pol and SNP PolTaq DNA polymerases differ up to 100% by means of allele-specific PCR between the two alleles and, after a simple qPCR, give a clear result as to which allele is present. Thus, the principly great potential of the CRISPR/Cas9 technology can be used for human medicine and plant biotechnology especially together with the SNP PolTaq or SNP Pol DNA polymerase. Allele-specific PCR can be used to quantify the mutation rate in a pool or background of wild-type sequences. The verification of mutation frequencies determined by NGS can also be verified by means of allele-specific PCR and SNP Pol DNA polymerase. The SNP PolTaq DNA polymerase is very suitable for the analysis of liquid biopsy samples. With SNP PolTaq, the presence and frequency of cancer mutations can be analyzed and quantified very well. Picture below: Application note SNP Pol DNA Polymerase We recommend short amplicon length (about 60-200 bp) for best results, but also longer amplicon lengths are possible. The addition of additional Magnesium (+0.5 - 1.5mM) might be needed in case of longer amplicons >500 bp.Trouble shooting: No bands after PCR of 30 cycles! Optimisation procedure for missing or weak bands.Annealing temperature is too low! Attention: The SNP Pol is an aptamer inhibited DNA polymerase. The aptamer oligo used reversibly inhibits the polymerase at temperatures <55°C! Therefore, the primers should ideally have an annealing temperature of >57°C. - realtime PCR approach - Check the dNTP concentration. This should be between 200 and 300µM (in the PCR). - Increase the number of cycles (at least 35, preferably equal to 40) Test optimisation - number of cyclesIn endpoint detection, a cycle count of 30 is not always sufficient to generate a clearly visible band. For example, with less than 200 DNA copies as a starting value. The test should therefore be run using real-time PCR, or five parallel PCRs should be used, one of which is taken from the PCR device after each of five different cycles (example below). Endpoint detection: Set up five identical PCR reactions in parallel using the SNP Pol DNA polymerase. Remove one of the PCR tubes from the cycler at each of 20, 25, 30, 35 and 40 cycles and apply all five reactions to an agarose gel at the end. This makes it easy to recognise which is the optimum number of cycles in the PCR for the given application (starting material, target, primer). SNP Pol PCR with cell lysates Animal cells and E.coli can be used directly for PCR with the SNP Pol DNA polymerase! No separate lysis or proteinase K digestion is necessary. PCR procedure:1. 50 - 500 cells are required per PCR batch! 2. prepare PCR mix with primers and buffer and place on ice! 3. add the picked colony directly to 10-20µL water (no separate lysis and no proteinase K digestion), vortex briefly (5 seconds), then add this cell suspension directly to the PCR reaction mixture, e.g. 10µL cell suspension for a PCR preparation with a total volume of 20µL. An initial denaturation time of 2-3 minutes is sufficient is sufficient, can be extended to 5 minutes if necessary. Genomic DNA per reaction max. 100 copies is relatively low, but should still work. The remainder of this cell suspension can also be frozen away in order to carry out further tests. Optional: 4. if possible: do real-time PCR!The SNP Pol DNA polymerase (M3009 > or M3061 >) can be used together with non-specific fluorescent dyes (eg Genaxxon's Green DNA Dye > or SybrGreen®) in real-time PCR. When working with specific PCR probes, only the SNP PolTaq DNA polymerase can be used, since only these have a 5'-3 'exonuclease activity. With our high quality dNTPs as Set (M3015.4100) > or Mix (M3016.1010) > or our DNALadders > and our favourable standard agarose (M3044) > we can offer additional products for your PCR. Application areas for SNP Pol DNA and SNP Pol DNA polymerase- Monitoring, verification and detection of point mutations- Identification of correct or wrong CRISPR/Cas9 products- Verification/validation of sequencing results- Quantification of mutations (e.g. NGS results)- SNP-detection by allele-specific amplification (ASA) / Allele-specific PCR- Methylation specific PCRs (MSP) after bisulfite treated DNA (CpG methylation sides)- HLA genotyping- micro sequencing- realtime PCR with hydrolysis probes- realtime multiplex PCRs - DamID-seq data in C. elegans. As an alternative to ChIP, DNA adenine methyltransferase identification by sequencing (DamID-seq) was recently shown to be able to characterize binding sites in single mammalian cells. Additionally, DamID can be achieved for cell-type specific analysis by expressing Dam fusion proteins under tissue specific promoters in a controlled manner. In this report, we present a user-friendly pipeline to analyse DamID-seq data in C. elegans. Sharma R, Ritler D, Meister P.Tools for DNA adenine methyltransferase identification analysis of nuclear organization during C. elegans development. Genesis. 2016 Feb 4. doi: 10.1002/dvg.22925. - Minisequencing SNP genotyping with SNPase DNA Polymerase can be carried out by the procedure described in: Lovmar L, Fredriksson M, Liljedahl U, Sigurdsson S, Syvänen AC. Quantitative evaluation by minisequencing and microarrays reveals accurate multiplexed SNP genotyping of whole genome amplified DNA. Nucleic Acids Res. 2003;31:e129. - Allel specific mismatch selectivity by the HiDi DNA polymerase Drum M, Kranaster R, Ewald C, Blasczyk R, Marx A (2014) Variants of a Thermus aquaticus DNA Polymerase with Increased Selectivity for Applications in Allele- and Methylation-Specific Amplification. PLoS ONE 9(5): e96640. doi:10.1371/journal.pone.0096640
The SNP Pol DNA polymerase > used for the SNP Pol 2X Mastermix has been specially designed for easy, reliable and rapid allele-specific discrimination, eg. CRISPR / Cas9 point mutations, for detecting incorrect CRISPR / Cas9 products, or validating sequencing results. The SNP Pol DNA polymerase distinguishes highly specific, whether a mismatch of the primer-template-complex is present or not. The mismatch (point mutation) must be at the 3 'end of the primer. Thus, mutant alleles can be distinguished exactly from wild-type alleles - without sequencing, since the polymerase simply does not amplify in the case of a mismatch. Just place your primer on the supposed point mutation (Important: The point mutation must be at the 3 'end) and the polymerase will detect a mismatch in this region with almost 100% accuracy: If the template base complementary to the 3' end of the primer shows the mutation and the primer does not, no amplification takes place - 100% certainty within a short time!The SNP Pol DNA polymerase can therefore be easily, time and cost-effectively used for the screening of point mutations. The variant SNP PolTaq DNA polymerase has 5'-3 'nuclease activity and can therefore be used for specific hydrolysi probes such as Taqman® probes or Molecular beacons. For more information on our SNP DNA polymerases and applications, read our blog now. DescriptionSNP Pol DNA Polymerase and SNP PolTaq Polymerase are highly selective DNA polymerase for the detection of Single Nucleotide Polymorphisms. It was developed specifically for allele-specific discrimination, where a very high discrimination rate is required e.g., in allele-specific PCR (ASA; AS-PCR), allele-specific primer extension (AS-PEX), SNP analysis, genotyping or in methylation-specific PCRs (MSP). Many other DNA polymerases tolerate mismatched primer-template complexes and are therefor not suitable. The SNP Pol DNA polymerase, on the other hand, distinguishes these specifically (high discrimination) and supplies only PCR products with perfectly matching primer pairs! The Genaxxon SNP Pol and SNP PolTaq DNA polymerases differ up to 100% by means of allele-specific PCR between the two alleles and, after a simple qPCR, give a clear result as to which allele is present. Thus, the principly great potential of the CRISPR/Cas9 technology can be used for human medicine and plant biotechnology especially together with the SNP PolTaq or SNP Pol DNA polymerase. Allele-specific PCR can be used to quantify the mutation rate in a pool or background of wild-type sequences. The verification of mutation frequencies determined by NGS can also be verified by means of allele-specific PCR and SNP Pol DNA polymerase. The SNP PolTaq DNA polymerase is very suitable for the analysis of liquid biopsy samples. With SNP PolTaq, the presence and frequency of cancer mutations can be analyzed and quantified very well. Picture below: Application note SNP Pol DNA Polymerase We recommend short amplicon length (about 60-200 bp) for best results, but also longer amplicon lengths are possible. The addition of additional Magnesium (+0.5 - 1.5mM) might be needed in case of longer amplicons >500 bp.Trouble shooting: No bands after PCR of 30 cycles! Optimisation procedure for missing or weak bands.Annealing temperature is too low! Attention: The SNP Pol is an aptamer inhibited DNA polymerase. The aptamer oligo used reversibly inhibits the polymerase at temperatures <55°C! Therefore, the primers should ideally have an annealing temperature of >57°C. - realtime PCR approach - Check the dNTP concentration. This should be between 200 and 300µM (in the PCR). - Increase the number of cycles (at least 35, preferably equal to 40) Test optimisation - number of cyclesIn endpoint detection, a cycle count of 30 is not always sufficient to generate a clearly visible band. For example, with less than 200 DNA copies as a starting value. The test should therefore be run using real-time PCR, or five parallel PCRs should be used, one of which is taken from the PCR device after each of five different cycles (example below). Endpoint detection: Set up five identical PCR reactions in parallel using the SNP Pol DNA polymerase. Remove one of the PCR tubes from the cycler at each of 20, 25, 30, 35 and 40 cycles and apply all five reactions to an agarose gel at the end. This makes it easy to recognise which is the optimum number of cycles in the PCR for the given application (starting material, target, primer). SNP Pol PCR with cell lysates Animal cells and E.coli can be used directly for PCR with the SNP Pol DNA polymerase! No separate lysis or proteinase K digestion is necessary. PCR procedure:1. 50 - 500 cells are required per PCR batch! 2. prepare PCR mix with primers and buffer and place on ice! 3. add the picked colony directly to 10-20µL water (no separate lysis and no proteinase K digestion), vortex briefly (5 seconds), then add this cell suspension directly to the PCR reaction mixture, e.g. 10µL cell suspension for a PCR preparation with a total volume of 20µL. An initial denaturation time of 2-3 minutes is sufficient is sufficient, can be extended to 5 minutes if necessary. Genomic DNA per reaction max. 100 copies is relatively low, but should still work. The remainder of this cell suspension can also be frozen away in order to carry out further tests. Optional: 4. if possible: do real-time PCR!The SNP Pol DNA polymerase (M3009 > or M3061 >) can be used together with non-specific fluorescent dyes (eg Genaxxon's Green DNA Dye > or SybrGreen®) in real-time PCR. When working with specific PCR probes, only the SNP PolTaq DNA polymerase can be used, since only these have a 5'-3 'exonuclease activity. With our high quality dNTPs as Set (M3015.4100) > or Mix (M3016.1010) > or our DNALadders > and our favourable standard agarose (M3044) > we can offer additional products for your PCR. Application areas for SNP Pol DNA and SNP Pol DNA polymerase- Monitoring, verification and detection of point mutations- Identification of correct or wrong CRISPR/Cas9 products- Verification/validation of sequencing results- Quantification of mutations (e.g. NGS results)- SNP-detection by allele-specific amplification (ASA) / Allele-specific PCR- Methylation specific PCRs (MSP) after bisulfite treated DNA (CpG methylation sides)- HLA genotyping- micro sequencing- realtime PCR with hydrolysis probes- realtime multiplex PCRs - DamID-seq data in C. elegans. As an alternative to ChIP, DNA adenine methyltransferase identification by sequencing (DamID-seq) was recently shown to be able to characterize binding sites in single mammalian cells. Additionally, DamID can be achieved for cell-type specific analysis by expressing Dam fusion proteins under tissue specific promoters in a controlled manner. In this report, we present a user-friendly pipeline to analyse DamID-seq data in C. elegans. Sharma R, Ritler D, Meister P.Tools for DNA adenine methyltransferase identification analysis of nuclear organization during C. elegans development. Genesis. 2016 Feb 4. doi: 10.1002/dvg.22925. - Minisequencing SNP genotyping with SNPase DNA Polymerase can be carried out by the procedure described in: Lovmar L, Fredriksson M, Liljedahl U, Sigurdsson S, Syvänen AC. Quantitative evaluation by minisequencing and microarrays reveals accurate multiplexed SNP genotyping of whole genome amplified DNA. Nucleic Acids Res. 2003;31:e129. - Allel specific mismatch selectivity by the HiDi DNA polymerase Drum M, Kranaster R, Ewald C, Blasczyk R, Marx A (2014) Variants of a Thermus aquaticus DNA Polymerase with Increased Selectivity for Applications in Allele- and Methylation-Specific Amplification. PLoS ONE 9(5): e96640. doi:10.1371/journal.pone.0096640
SNP Pol DNA polymerase for easy, reliable and rapid allele-specific discrimination, eg. CRISPR / Cas9 point mutations, for detecting incorrect CRISPR / Cas9 products, or validating sequencing results. The SNP Pol DNA polymerase distinguishes highly specific, whether a mismatch of the primer template complex is present or not. The mismatch (point mutation) must be at the 3 'end of the primer. Thus, mutant alleles can be distinguished exactly from wild-type alleles - without sequencing since the polymerase simply does not amplify in the case of a mismatch. Just place your primer on the supposed point mutation (Important: The point mutation must be at the 3 'end) and the polymerase will detect a mismatch in this region with almost 100% accuracy: If the template base complementary to the 3' end of the primer shows the mutation and the primer does not, no amplification takes place - 100% certainty within a short time!The SNP Pol DNA polymerase can therefore be easily, time and cost-effectively used for the screening of point mutations. SNP PolTaq DNA polymerase shows also 5'-3'-nuclease activity and is therefore suitable for hydrolysis probe-based assays (Taqman®, molecular beacons, etc.). For more information on our SNP DNA polymerases and applications, read our blog now. DescriptionSNP PolTaq DNA Polymerase is a highly selective DNA polymerase for the detection of Single Nucleotide Polymorphisms. It was developed specifically for allele-specific discrimination, where a very high discrimination rate is required e.g., in allele-specific PCR (ASA; AS-PCR), allele-specific primer extension (AS-PEX), SNP analysis, genotyping or in methylation-specific PCRs (MSP). Many other DNA polymerases tolerate mismatched primer-template complexes and are therefor not suitable. The SNP Pol DNA polymerase, on the other hand, distinguishes these specifically (high discrimination) and supplies only PCR products with perfectly matching primer pairs! The Genaxxon SNP Pol and SNP PolTaq DNA polymerases differ up to 100% by means of allele-specific PCR between the two alleles and, after a simple qPCR, give a clear result as to which allele is present. Thus, the principle great potential of the CRISPR/Cas9 technology can be used for human medicine and plant biotechnology especially together with the SNP PolTaq or SNP Pol DNA polymerase. Allele-specific PCR can be used to quantify the mutation rate in a pool or background of wild-type sequences. The verification of mutation frequencies determined by NGS can also be verified by means of allele-specific PCR and SNP Pol DNA polymerase. The SNP PolTaq DNA polymerase is very suitable for the analysis of liquid biopsy samples. With SNP PolTaq , the presence and frequency of cancer mutations can be analyzed and quantified very well. Picture below: Application note SNP PolTaq DNA Polymerase We recommend short amplicon length (about 60-200 bp) for best results, but also longer amplicon lengths are possible. The addition of additional Magnesium (+0.5 - 1.5mM) might be needed in case of longer amplicons >500 bp.Trouble shooting: No bands after PCR of 30 cycles! Optimisation procedure for missing or weak bands.Annealing temperature is too low! Attention: The SNP PolTaq is an aptamer inhibited DNA polymerase. The aptamer oligo used reversibly inhibits the polymerase at temperatures <55°C! Therefore, the primers should ideally have an annealing temperature of >57°C. - realtime PCR approach - Check the dNTP concentration. This should be between 200 and 300µM (in the PCR). - Increase the number of cycles (at least 35, preferably equal to 40) Test optimisation - number of cyclesIn endpoint detection, a cycle count of 30 is not always sufficient to generate a clearly visible band. For example, with less than 200 DNA copies as a starting value. The test should therefore be run using real-time PCR, or five parallel PCRs should be used, one of which is taken from the PCR device after each of five different cycles (example below). Endpoint detection: Set up five identical PCR reactions in parallel using the SNP Pol DNA polymerase. Remove one of the PCR tubes from the cycler at each of 20, 25, 30, 35 and 40 cycles and apply all five reactions to an agarose gel at the end. This makes it easy to recognise which is the optimum number of cycles in the PCR for the given application (starting material, target, primer). SNP Pol PCR with cell lysates Animal cells and E.coli can be used directly for PCR with the SNP Pol DNA polymerase! No separate lysis or proteinase K digestion is necessary. PCR procedure:1. 50 - 500 cells are required per PCR batch! 2. prepare PCR mix with primers and buffer and place on ice! 3. add the picked colony directly to 10-20µL water (no separate lysis and no proteinase K digestion), vortex briefly (5 seconds), then add this cell suspension directly to the PCR reaction mixture, e.g. 10µL cell suspension for a PCR preparation with a total volume of 20µL. An initial denaturation time of 2-3 minutes is sufficient is sufficient, can be extended to 5 minutes if necessary. Genomic DNA per reaction max. 100 copies is relatively low, but should still work. The remainder of this cell suspension can also be frozen away in order to carry out further tests. Optional: 4. if possible: do real-time PCR!The SNP Pol DNA polymerase (M3009 > or M3061 >) can be used together with non-specific fluorescent dyes (eg Genaxxon's Green DNA Dye > or SybrGreen®) in real-time PCR. When working with specific PCR probes, only the SNP PolTaq DNA polymerase can be used, since only these have a 5'-3 'exonuclease activity. With our high quality dNTPs as Set (M3015.4100) > or Mix (M3016.1010) > or our DNALadders > and our favourable standard agarose (M3044) > we can offer additional products for your PCR. Application areas for SNP Pol DNA and SNP PolTaq DNA polymerase- Monitoring, verification and detection of point mutations- Identification of correct or wrong CRISPR/Cas9 products- Verification/validation of sequencing results- Quantification of mutations (e.g. NGS results)- SNP-detection by allele-specific amplification (ASA) / Allele-specific PCR- Methylation specific PCRs (MSP) after bisulfite treated DNA (CpG methylation sides)- HLA genotyping- micro sequencing- realtime PCR with hydrolysis probes- realtime multiplex PCRs - DamID-seq data in C. elegans. As an alternative to ChIP, DNA adenine methyltransferase identification by sequencing (DamID-seq) was recently shown to be able to characterize binding sites in single mammalian cells. Additionally, DamID can be achieved for cell-type specific analysis by expressing Dam fusion proteins under tissue specific promoters in a controlled manner. In this report, we present a user-friendly pipeline to analyse DamID-seq data in C. elegans. Sharma R, Ritler D, Meister P.Tools for DNA adenine methyltransferase identification analysis of nuclear organization during C. elegans development. Genesis. 2016 Feb 4. doi: 10.1002/dvg.22925. - Minisequencing SNP genotyping with SNPase DNA Polymerase can be carried out by the procedure described in: Lovmar L, Fredriksson M, Liljedahl U, Sigurdsson S, Syvänen AC.bQuantitative evaluation by minisequencing and microarrays reveals accurate multiplexed SNP genotyping of whole genome amplified DNA. Nucleic Acids Res. 2003;31:e129. - Allel specific mismatch selectivity by the HiDi DNA polymerase Drum M, Kranaster R, Ewald C, Blasczyk R, Marx A (2014) Variants of a Thermus aquaticus DNA Polymerase with Increased Selectivity for Applications in Allele- and Methylation-Specific Amplification. PLoS ONE 9(5): e96640. doi:10.1371/journal.pone.0096640
The SNP PolTaq DNA polymerase used for the SNP PolTaq 2X Master Mix has been specially designed for easy, reliable and rapid allele-specific discrimination, eg. CRISPR / Cas9 point mutations, for detecting incorrect CRISPR / Cas9 products, or validating sequencing results. The SNP PolTaq DNA polymerase distinguishes highly specific, whether a mismatch of the primer-template-complex is present or not. The mismatch (point mutation) must be at the 3 'end of the primer. Thus, mutant alleles can be distinguished exactly from wild-type alleles - without sequencing, since the polymerase simply does not amplify in the case of a mismatch. Just place your primer on the supposed point mutation (Important: The point mutation must be at the 3 'end) and the polymerase will detect a mismatch in this region with almost 100% accuracy: If the template base complementary to the 3' end of the primer shows the mutation and the primer does not, no amplification takes place - 100% certainty within a short time!The SNP PolTaq DNA polymerase can therefore be easily, time and cost-effectively used for the screening of point mutations. The SNP PolTaq DNA polymerase has 5'-3 'nuclease activity and can therefore be used for specific hydrolysi probes such as Taqman® probes or Molecular beacons. For more information on our SNP DNA polymerases and applications, read our blog now. DescriptionSNP PolTaq DNA polymerase is a highly selective DNA polymerase for the detection of Single Nucleotide Polymorphisms. It was developed specifically for allele-specific discrimination, where a very high discrimination rate is required e.g., in allele-specific PCR (ASA; AS-PCR), allele-specific primer extension (AS-PEX), SNP analysis, genotyping or in methylation-specific PCRs (MSP). Many other DNA polymerases tolerate mismatched primer-template complexes and are therefor not suitable. The SNP PolTaq DNA polymerase, on the other hand, distinguishes these specifically (high discrimination) and supplies only PCR products with perfectly matching primer pairs! The Genaxxon SNP Pol and SNP PolTaq DNA polymerases differ up to 100% by means of allele-specific PCR between the two alleles and, after a simple qPCR, give a clear result as to which allele is present. Thus, the principly great potential of the CRISPR/Cas9 technology can be used for human medicine and plant biotechnology especially together with the SNP PolTaq or SNP Pol DNA polymerase. Allele-specific PCR can be used to quantify the mutation rate in a pool or background of wild-type sequences. The verification of mutation frequencies determined by NGS can also be verified by means of allele-specific PCR and SNP Pol DNA polymerase. The SNP PolTaq DNA polymerase is very suitable for the analysis of liquid biopsy samples. With SNP PolTaq, the presence and frequency of cancer mutations can be analyzed and quantified very well. Picture below: Application note SNP Pol DNA Polymerase We recommend short amplicon length (about 60-200 bp) for best results, but also longer amplicon lengths are possible. The addition of additional Magnesium (+0.5 - 1.5mM) might be needed in case of longer amplicons >500 bp.Trouble shooting: No bands after PCR of 30 cycles! Optimisation procedure for missing or weak bands.Annealing temperature is too low! Attention: The SNP PolTaq is an aptamer inhibited DNA polymerase. The aptamer oligo used reversibly inhibits the polymerase at temperatures <55°C! Therefore, the primers should ideally have an annealing temperature of >57°C. - realtime PCR approach - Check the dNTP concentration. This should be between 200 and 300µM (in the PCR). - Increase the number of cycles (at least 35, preferably equal to 40) Test optimisation - number of cyclesIn endpoint detection, a cycle count of 30 is not always sufficient to generate a clearly visible band. For example, with less than 200 DNA copies as a starting value. The test should therefore be run using real-time PCR, or five parallel PCRs should be used, one of which is taken from the PCR device after each of five different cycles (example below). Endpoint detection: Set up five identical PCR reactions in parallel using the SNP Pol DNA polymerase. Remove one of the PCR tubes from the cycler at each of 20, 25, 30, 35 and 40 cycles and apply all five reactions to an agarose gel at the end. This makes it easy to recognise which is the optimum number of cycles in the PCR for the given application (starting material, target, primer). SNP Pol PCR with cell lysates Animal cells and E.coli can be used directly for PCR with the SNP Pol DNA polymerase! No separate lysis or proteinase K digestion is necessary. PCR procedure:1. 50 - 500 cells are required per PCR batch! 2. prepare PCR mix with primers and buffer and place on ice! 3. add the picked colony directly to 10-20µL water (no separate lysis and no proteinase K digestion), vortex briefly (5 seconds), then add this cell suspension directly to the PCR reaction mixture, e.g. 10µL cell suspension for a PCR preparation with a total volume of 20µL. An initial denaturation time of 2-3 minutes is sufficient is sufficient, can be extended to 5 minutes if necessary. Genomic DNA per reaction max. 100 copies is relatively low, but should still work. The remainder of this cell suspension can also be frozen away in order to carry out further tests. Optional: 4. if possible: do real-time PCR!The SNP Pol DNA polymerase (M3009 > or M3061 >) can be used together with non-specific fluorescent dyes (eg Genaxxon's Green DNA Dye > or SybrGreen®) in real-time PCR. When working with specific PCR probes, only the SNP PolTaq DNA polymerase can be used, since only these have a 5'-3 'exonuclease activity. With our high quality dNTPs as Set (M3015.4100) > or Mix (M3016.1010) > or our DNALadders > and our favourable standard agarose (M3044) > we can offer additional products for your PCR. Application areas for SNP Pol DNA and SNP PolTaq DNA polymerase- Monitoring, verification and detection of point mutations- Identification of correct or wrong CRISPR/Cas9 products- Verification/validation of sequencing results- Quantification of mutations (e.g. NGS results)- SNP-detection by allele-specific amplification (ASA) / Allele-specific PCR- Methylation specific PCRs (MSP) after bisulfite treated DNA (CpG methylation sides)- HLA genotyping- micro sequencing- realtime PCR with hydrolysis probes- realtime multiplex PCRs - DamID-seq data in C. elegans. As an alternative to ChIP, DNA adenine methyltransferase identification by sequencing (DamID-seq) was recently shown to be able to characterize binding sites in single mammalian cells. Additionally, DamID can be achieved for cell-type specific analysis by expressing Dam fusion proteins under tissue specific promoters in a controlled manner. In this report, we present a user-friendly pipeline to analyse DamID-seq data in C. elegans. Sharma R, Ritler D, Meister P.Tools for DNA adenine methyltransferase identification analysis of nuclear organization during C. elegans development. Genesis. 2016 Feb 4. doi: 10.1002/dvg.22925. - Minisequencing SNP genotyping with SNPase DNA Polymerase can be carried out by the procedure described in: Lovmar L, Fredriksson M, Liljedahl U, Sigurdsson S, Syvänen AC. Quantitative evaluation by minisequencing and microarrays reveals accurate multiplexed SNP genotyping of whole genome amplified DNA. Nucleic Acids Res. 2003;31:e129. - Allel specific mismatch selectivity by the HiDi DNA polymerase Drum M, Kranaster R, Ewald C, Blasczyk R, Marx A (2014) Variants of a Thermus aquaticus DNA Polymerase with Increased Selectivity for Applications in Allele- and Methylation-Specific Amplification. PLoS ONE 9(5): e96640. doi:10.1371/journal.pone.0096640