Advantages at a glance
- High-quality products
- Customized solutions
- Personal contact
- Fast service
- Competent technical support +49(0)731-3608-123
- Free shipping
- Shipping within 24h
- 30 days money back guarantee
- Purchase comfortable on account
Product information "SuperHot Taq PCR Kit with dNTP mix"
SuperHot Taq DNA polymerase is a chemicaly modified form of thermostable DNA polymerase, which is activated by heat treatment. A chemical moiety is attached to the enzyme at the active site, which renders the enzyme inactive at room temperature. Thus, during setup and the first PCR cycle, the enzyme is not active and misprimed primers are not extended. As a result specifity and efficiency are increased by far compared to standard Taq DNA polymerase. Additionally, difficult targets with high GC-content can be amplified. Sensitivity improves multiplex PCR, an applied PCR technique that amplifies several specific targets simultaneously. The Genaxxon SuperHot Taq-Kit includes Genaxxon's SuperHot Taq Polymerase, buffer E, MgCl2, appropriate amounts of a 10mM dNTP-Mix sufficient for up to 500 / 2000 x 25µL PCR reactions (0.5 units Taq per reaction). All items are in shipped in separate tubes.
With our high quality dNTPs as Set (M3015.4100 and M3015.0250) > or Mix (M3016.1010) > or our DNA Ladders > and favourable standard agarose (M3044) > we can offer additional products for your PCR.
Specifications:
HotStart Taq polymerasee with 5 units/µL. Kit contains also 100µL of a 10mM dNTP mix.
Enyzme is completely inactive prior to heating at 95°C for at least 10 min.!
Application:
For Real-time PCR, for PCR of low amount of DNA (very low copy number).Units:
One unit is defined as the amount of enzyme which will convert 10 nmoles of dNTPs to an acid-insoluble form in 30 min at 72°C under the assay conditions (25 mM TAPS (tris-(hydroxymethyl)-methyl-amino-propanesulfonic acid, sodium salt) pH 9.3 (25°C); 50 mM KCl; 2 mM MgCl2; 1 mM b-mercaptoethanol; and activated calf thymus DNA as substrate.Source:
recombinantClassification:
Documents:
Safety Data SheetProtocols
Certificate
Product description
General Data 3
Category List
DNA Template
Amplification of templates with high GC content, strong secondary structure, low concentrations, or which produce products greater than 5 kb, may require adaptation of the following parameters:
- Use high quality, purified DNA templates.
- Approximately 10E4 copies of target DNA are required to detect product in 25-30 PCR cycles.
- Use 1pg–1ng of plasmid or viral templates.
- Use 1ng–1µg of genomic templates.
- Higher DNA concentrations decrease amplicon specificity (i.e., extra bands are more likely), particularly when a large number of cycles are employed.
- Use the higher DNA concentrations when fewer cycles are desired (e.g. to increase fidelity).
Primers
- Generally 20-30 nucleotides in length.
- Ideal GC content is 40-60%.
- Space GC residues evenly within the primer.
- Primer pairs should have Tms within 5°C of each other.
- Avoid secondary structure (i.e., hairpins) within each primer and potential dimerization between the primers present.
- When engineering sites into the end of primers, 4-6 extra bases should be added 5´ to the site.
- Final concentration should be 0.05-1 µM, typically 0.1-0.5 µM of each primer.
- Higher concentrations may increase secondary priming and create spurious amplification products.
Magnesium Concentration
- 1.5-2.0 mM is optimal for Taq DNA Polymerase, but the ideal concentration depends on template, buffer, DNA and dNTPs (each has the potential to chelate magnesium).
- If [Mg2+] is too low, no PCR product will be seen.
- If [Mg2+] is too high, undesired PCR products may be seen.
Deoxyribonucleotide triphosphates (dNTPs) >
- Typical concentration is 200 µM of each dNTP.
- 50-100 µM enhances fidelity of polymerization, but reduces yields.
- Higher concentrations increase yields particularly in long PCR, but can reduce fidelity.
DNA Polymerase
- The choice of the correct polymerase depends among other things on the purpose as well as on the template used (standard PCR: Taq DNA Polymerase S > with high accuracy, Taq Polymerase E > with high yield; master mixes: Standard PCR master mix > or RedMasterMix > with red dye).
- For multiplex PCR, there exist special multiplex master mixes >.
- Hot start applications are recommended to increase specificity or if you use difficult templates. For this purpose there exist special Hot Start Polymerases M3307 > or M3006 >.
- For PCRs for the purpose of cloning or other procedures requiring a low error rate, there are thermostable high fidelity proofing polymerases such as Pfu >, AQ97>, or ReproFast >. These enzymes make far fewer errors during amplification and increase the chances of an amplicon without mistakes.
- For genotyping and other applications where a high discrimination rate is required, there is a new highly selective DNA polymerase, SNP Pol DNA Polymerase >. It specifically distinguishes mismatched primer-template complexes and ony produces specific amplicons in case of perfectly matched primer pairs.
- For the real-time quantitative PCR there exist highly specialized qPCR master mixes >. Hereby, the choice of the correct master mix depends on the qPCR device you are using. Alternatively, you can simply use a universal master mix, such as the GNX Versa qPCR MasterMix 2X > from Genaxxon, which automatically ‘adjusts’ the correct amount of ROX for the respective instrument. Even if there are different PCR cyclers in the laboratory, you no longer need to worry about which master mix to use
- The amount of DNA polymerase used in the PCR reaction can significantly influence the PCR result (use 1.25-1.5 units Taq Polymerase > for a 50μL volume).
General Guidelines
Annealing Temperature and Duration
- Match the Tms within 5°C of each other.
- Typical annealing temperatures are 5°C below the lowest primer's Tm and often fall in the range of 50-60°C.
- Test higher annealing temperatures if spurious amplification products are observed.
- Typical annealing times are 15-30 seconds.
Extension Time
- Extensions are normally performed at 68°C.
- In general, use extension times of one minute per 1000 base (1 kb) pairs (e.g. 3 minutes for a 3 kb product).
- For products less than 1 kb, use 45-60 seconds.
- Products greater than 3 kb, or reactions using more than 30 cycles, may require longer extensions.
The amplification of templates with high GC content, strong secondary structures, low concentrations or amplicons more than 5 kb often require optimization of the PCR conditions. Typically, 15-30 seconds of denaturation should be performed at 95 ° C during PCR.
Source: NCBI PubMed
Accessory Items
Our DNA marker GenLadder 100 bp Plus with loading dye is an ideal tool to determine the size of double stranded DNA in the range of 100 - 3000 bp. The additional bands at 1.5kbp and 3.0kbp allow to assign additional DNA fragment sizes in the range up to 3000 bp without using an additional 1kbp DNA marker. The Gene Ladder 100 bp Plus consists of 12 fragments in sizes between 100 - 1000bp spaced 100bp apart. In addition, there are further bands at 1500bp and 3000bp. The 500bp and 1500bp fragment show stronger intensity to allow easier identification. All fragments are 'blunt-ended'. The GenLadder can be directly labeled at the 5'-end with radioisotopes using T4 polynucleotide Kinase for visualisation by autoradiography. Fragment sizes (in base pairs): 3000 bp (40ng/5µL), 2x 1500 bp (70ng/5µL), 1000 bp (50ng/5µL), 900 bp (40ng/5µL), 800 bp (40ng/5µL), 700 bp (30ng/5µL), 600 bp (30ng/5µL), 2x 500 bp (90ng/5µL), 400 bp (40ng/5µL), 300 bp (30ng/5µL), 200 bp (40ng/5µL), 100 bp (40ng/5µL). The DNA marker is already dissolved in buffer containing orange G and Xylene cyanol FF as tracking dyes. The total concentration of the DNA-Marker is 0.1µg/µL (108µg/mL). The recommended amount of DNA marker per lane is 0,5μg (5µL). GenLadder DNA ladders are available in a ready-to-use format (premixed with 6X DNA Loading Dye): M3094 (GenLadder 1kbp extended up to 25 kbp ready-to-use) > and M3328 (GenLadder 1kbp ready-to-use) > or our ready-to-use premix M3072 (GenLadder 50bp) > with 6X Orange DNA Loading Dye. The DNA Gel Loading Dyes can also be ordered separately: M3308 (DNA Gel Loading Dye with Bromophenol and Xylene Cyanol FF) >, M3321 (DNA Gel Loading Dye with Orange G) >.
Agarose LE is a standard agarose for the separation of DNA in the size range between 100bp and 25kbp. It is suitable for all analytical and preparative electrophoresis of nucleic acids in routine gel electrophoresis. Depending on the concentration of Agarose LE used, the size range of nucleic acid separation will vary between 100bp and 25kbp. The low EEO makes this useful for a broad range of applications: PCR product analysis, restriction enzyme digest analysis, separation of RNA before blotting, etc. This agarose is comparable with, e.g. Agarose BioRagent, low EEO from Sigma or with the Universal-Agarose, peqGOLD from Peqlab. Free Taq DNA Polymerase test sample available! No shipping costs within Germany. Genaxxon offers Agaroses for different purposes. Two melting point options and normal or high resolution separation of DNA fragments are available. Standard normal melting agarose M3044 with standard resolution used in routine DNA electrophoresis for separation of a wide range of DNA fragments from 100bp up to 25kbp. This agarose is available as powder (M3044 Agarose LE >) or as high-quality, multi-purpose tablets (M3054 >) available in blister packaging in 200 or 1,000 tablet quantities. They are compact, pre-weight and therefore easy-to-use. Low melting/gelling temperature agarose recommended for rapid DNA gel extraction with the agarose-digesting enzyme, agarase, as well as for "in-gel" DNA treatment with enzymes or for bacterial transformation with nucleic acids directly after re-melting the gel. High resolution agarose (normal melting or low melting) for high resolution elektrophoresis to differentiate DNA fragments with only 2 bp size difference: M3046 Agarose Tiny > (low melt, high resolution comparable to NuSieve®), or M3047 Agarose Tiny HT > (high gel strength, high resolution comparable to SeaKem®). For the nontoxic staining of nucleic acids in agarose gels the highly sensitive fluorescent dye SafeGel red stain (M3193) or SafeGel green stain (M3191) is recommended.
Highly pure, HPLC purified (>99%) dNTPs packaged as 4 separate 100mM solutions of dATP, dCTP, dGTP and dTTP for qPCR, RT-PCR, standard PCR, and Klenow reactions. Genaxxon’s dNTP solutions have been optimized for use in DNA amplification and other related methods. Genaxxon dNTPs contain no measurable bacterial or human DNA. For long term storage and/or for repeated use, our recommendation is to aliquot the stock solutions. Solutions of dNTP sodium salts (dATP, dCTP, dGTP und dTTP) Concentration: 100 mM each nucleotide Please have also a look on our broad range of nucleotides especially the dNTP mixes with 2 mM or 10 mM or the modified nucleotides, e.g. Biotin-11-dUTP. 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.
The GenLadder 1kb (ready-to-use) 250bp - 10kbp-ladder is the ideal DNA size marker for fragment sizes from 250 - 10000bp. All bands light up to the same intensity except the 1000 bp and the 3000 bp band which are twice as bright for better orientation. Each band has been calibrated with respect to base pair size and DNA quantity. Thus, an accurate determination of product size and DNA concentration is possible. No ambiguous bands are visible. The GenLadder 250bp - 10kb can be directly labeled at the 5'-end with radioisotopes using T4 polynucleotide Kinase for visualisation by autoradiography. The DNA is already dissolved in buffer with 6X loading dye ready to use. Fragment sizes and amount of DNA per band: 10000bp ( 28ng/5µL), 8000bp (28ng/5µL), 6000bp (28ng/5µL), 5000bp (28ng/5µL), 4000bp ( 18ng/5µL), 2x3000bp (92ng/5µL), 2500bp (34ng/5µL), 2000bp (34ng/5µL), 1500bp (20ng/5µL), 2x1000bp (92ng/5µL), 750bp (23ng/5µL), 500bp (30ng/5µL), 250bp (45ng/5µL). GenLadder DNA ladders are available in a ready-to-use format (premixed with 6X DNA Loading Dye): M3094 (GenLadder 1kbp extended up to 25 kbp ready-to-use) > and M3328 (GenLadder 1kbp ready-to-use) > or our ready-to-use premix M3072 (GenLadder 50bp) > with 6X Orange DNA Loading Dye. The DNA Gel Loading Dyes can also be ordered separately: M3308 (DNA Gel Loading Dye with Bromophenol and Xylene Cyanol FF) >, M3321 (DNA Gel Loading Dye with Orange G) >.
Agarose Tiny is a low-melting high resolution agarose that is suitable for molecular biology applications and produces gels with better separation properties and greater clarity than standard agarose. It is very transparent even at high gel concentrations (3.0 - 4.5%) and can be used with all buffer systems. The low melting temperature makes Agarose Tiny ideal for preparative DNA and RNA electrophoresis, while the low gelling temperature is ideal for tissue culture cell cloning experiments and viral plaque assays. In addition to the above mentioned properties, Agarose Tiny can also be used for high-resolution (2 bp) gel electrophoresis in the range from 10 to 500 bp. Example: PCR products or small DNA fragments after restriction digestion or in mutation analyses. Features:Like Lonza's SeaPlaque™, Agarose Tiny has a low melting point (<64°C, 3.0% gel) and gel point (<25°C, 3.0% gel) and low DNA-binding capacity. Their use is particularly recommended for preparative RNA and DNA gels. Genaxxon offers Agaroses for different purposes. Two melting point options and normal or high resolution separation of DNA fragments are available. Standard agarose high melting point agarose M3044 with standard resolution used in routine DNA electrophoresis for separation of a wide range of DNA fragments from 50bp up to 20kbp. This agarose is available as powder (M3044 Agarose LE >) or as high-quality, multi-purpose tablets (M3054 >) available in blister packaging in 200 or 1,000 tablet quantities. These are compact, pre-weight and therefore easy-to-use. Low melting/gelling temperature agarose recommended for rapid DNA gel extraction with the agarose-digesting enzyme, Agarase, as well as for “in-gel’ DNA treatment with enzymes or for bacterial transformation with nucleic acids directly after re-melting the gel. High resolution agarose (normal melting or low melting) for high resolution elektrophoresis to differentiate DNA fragments with only 2 bp size difference: M3046 Agarose Tiny > (low melt, high resolution comparable to NuSieve®), or M3047 Agarose Tiny HT > (high gel strength, high resolution comparable to SeaKem®). For the nontoxic staining of nucleic acids in agarose gels the highly sensitive fluorescent dye SafeGel red stain (M3193) or SafeGel green stain (M3191) is recommended.Recommended for the following applications:- RFLP (Restriction Fragment Length Polymorphism - AMPFLP (Amplified Fragment Length Polymorphism) - HVR (Hyper-Variable Regionen) - VNTR (Variable Numbers of Tandem Repeats) - STR (Short Tandem Repeats) - Tri und Tetra Nukleotid repeats
Accessory Items
Highly specific Hotstart master mix for routine and diagnostic PCR and qPCR up to 4kb: ready-to-use master mix with chemical modified Taq DNA polymerase. This Hotstart Master Mix for qPCR is a premixed, ready-to-use solution containing chemically modified Taq DNA Polymerase, dNTPs, MgCl2 and reaction buffers at optimal concentrations for efficient amplification of DNA templates by PCR and qPCR (one tube, one pipetting step). Just add your primers and template DNA. The Hotstart PCR Master Mix has been optimized for use in routine and diagnostic Hotstart PCR and qPCR for amplifying DNA template in the range of 0.2-4kb. The special formulation of our Hotstart Master Mix guarantees you reproducible results even after repeated freezing/thawing steps leading to good yields and sensitivity. The Hotstart Master Mix is efficient (no left over of reagents), scalable from 10µL to 50µL and stable for 12 months. This Hotstart PCR master mix is also available as SuperHot Mastermix Blue > enabling visualisation of each pipetting step. With our high quality dNTPs as Set (M3015.4100 and M3015.0250) > or Mix (M3016.1010) > or our DNA Ladders > and favourable standard agarose (M3044) > we can offer additional products for your PCR.
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 >.
SuperHot Taq DNA Polymerase is a superior DNA Polymerase for Real Time PCR, Hot-Start PCR, low-copy number PCR, or PCR of difficult templates. The enzyme is developed to enhance the specificity, sensitivity and yield of DNA amplification. SuperHot Taq DNA polymerase is a chemicaly modified form of thermostable DNA polymerase Taq, which is activated by heat treatment. A chemical moiety is attached to the enzyme at the active site, which renders the enzyme inactive at room temperature. Thus, during setup and before the first PCR cycle, the enzyme is not active and misprimed primers are not extended. As a result specifity and efficiency are increased by far compared to standard Taq DNA polymerase. Additionally, difficult targets with high GC-content can be amplified. For realtime PCR Genaxxon offers specially optimized 2-times qPCR mastermix >, respective a special Multiplex-PCR mastermix >. Features of Genaxxons SuperHot DNA Polymerase chemically modified increased specifity increased sensitivity for difficult templates with high GC-content Please have also a look on our SuperHot Taq 2-times Mastermix >. Even more convenient than the SuperHot Taq. With our high quality dNTPs as Set (M3015.4100 and M3015.0250) > or Mix (M3016.1010) > or our DNA Ladders > and favourable standard agarose (M3044) > we can offer additional products for your PCR.
PCR Hotstart Master mix with blue dye: for the visual control of pipetting steps after PCR, electrophoresis can be operated without the addition of loading buffer. This makes this PCR master mix saving time and money. This Hotstart Master Mix for qPCR is a premixed, ready-to-use solution containing chemically modified Taq DNA Polymerase, dNTPs, MgCl2, a blue dye and reaction buffers at optimal concentrations for efficient amplification of DNA templates by PCR and qPCR (one tube, one pipetting step). Just add your primers and template DNA. The addition of the blue loading dye enables direct application of the PCR reaction onto a Gel. The Hotstart PCR Master Mix has been optimized for use in routine and diagnostic Hotstart PCR and qPCR for amplifying DNA template in the range of 0.2-4kb. The special formulation of our Hotstart Master Mix guarantees you reproducible results even after repeated freezing/thawing steps leading to good yields and sensitivity. The Hotstart Master Mix with blue dye is efficient (no left over of reagents), scalable from 10µL to 50µL and stable for 12 months. Hotstart Mastermix with blue dye for 200 x 25µL reactions (2.5mL). This Hotstart PCR master mix is also available as SuperHot Mastermix > enabling visualisation of each pipetting step. With our high quality dNTPs as Set (M3015.4100 and M3015.0250) > or Mix (M3016.1010) > or our DNA Ladders > and favourable standard agarose (M3044) > we can offer additional products for your PCR.
Taq DNA Hot Start Polymerase with Aptamer technology. The specific aptamer inhibits DNA polymerase activity below temperatures of 50°C to 55°C, thus enabling hot start PCR. At higher temperatures, the aptamer falls off the active site of Taq polymerase. The Hot Start Taq Polymerase can be used like a normal Taq without having to lengthen the first denaturation step.The binding of aptamer and polymerase prevents the formation of primer-dimer constructs and other artefacts revealed by non-specific primer binding. Check also our SuperHot Mastermix with or without dye. With our high quality dNTPs as Set or Mix or our DNA Ladders and our favourable standard agarose we can offer additional products for your PCR.