SNP PolTaq DNA Polymerase



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SNP Pol DNA polymerase for easy, reliable and rapid allele-specific discrimination, eg. CRISPR /... more
Product information "SNP PolTaq DNA Polymerase"

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.).

Test sample available at a special price. No shipment costs within Germany. The test sample price will be refunded on the first official order of the product.

Description
SNP Pol DNA Polymerase > (High Discrimination of single nucleotides) is a highly selective DNA polymerase. It has been specially developed for allele-specific discrimination where a high discrimination rate is needed: 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

Application Note for HiDi DNA Polymerase

 

We recommend designing primers with a 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.

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 and M3015.0250) > or Mix (M3016.1010) > or our DNA Ladders > and our favourable standard agarose (M3044) > Genaxxon 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.b
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

frequently asked questions (FAQs) >

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high single nucleotide discrimination
Specifications: SNP PolTaq DNA polymerase is supplied as a 5 U/µL solution. It comes together... more
 

Technical Data:

Specifications:
SNP PolTaq DNA polymerase is supplied as a 5 U/µL solution. It comes together with an optimized 10X reaction buffer.

SNP PolTaq DNA polymerase posses 5’-3’ exonuclease activity! This enzyme can be used together with hydrolysis probes like, e.g. TagMan probes.

application:

- 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 - HLA genotyping - micro sequencing - realtime PCR with hydrolysis probes - realtime multiplex PCRs

Source

rec. from E.coli

Sicherheits Hinweise / Safety

Klassifizierungen / Classification

eclass-Nr: 34-16-04-90
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Dokumente - Protokolle - Downloads

Here you will find information and further literature on SNP PolTaq DNA Polymerase. For further documents (certificates with additional lot numbers, safety data sheets in other languages, further product information) please contact Genaxxon biosience at: info@genaxxon.com or phone: +49 731 3608 123.

 
 
 
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Dokumente - Protokolle - Downloads

Kann man die SNP Pol DNA-Polymerase mit Bisulfit-behandelten DNA-Proben verwenden?

Ja, unsere SNP Pol DNA-Polymerase ist beispielsweise hervorragend geeignet für die methylierungsspezifische PCR (MSP). Da eine einzelne Fehlpaarung für bestimmte MSP-Ergebnisse ausreicht, ermöglicht SNP Pol DNA-Polymerase sogar eine zuverlässige MSP-Analyse einzelner CpG-Stellen.

Kann man SNP Pol 2x PCR Master Mix und SNP Pol DNA-Polymerase in auf Sonden basierenden Assays verwenden?

Ja, das ist möglich, solange keine Nukleaseaktivität erforderlich ist. Bitte beachten Sie, dass SNP Pol DNA-Polymerase keine 5'-3'-Nuklease-Aktivität aufweist. SNP Pol 2x-PCR Master Mix kann nicht in hydrolysebasierten Sondenassays (z. B. TaqMan®) verwendet werden. Für diese Assays empfehlen wir die Verwendung unserer SNP PolTaq DNA-Polymerase mit 5'-3'-Nuklease-Aktivität. Die SNP PolTaq-Variante kann daher für auf Hydrolysesonden-basierenden Real Time-PCRs verwendet werden.

Was ist die Fehlerrate von SNP Pol DNA-Polymerase?

Die Fehlerrate von SNP Pol ist vergleichbar mit der Fehlerrate von Wildtyp-Taq DNA-Polymerase.

Kann man SNP Pol DNA-Polymerase in Real Time-PCRs mit einem qPCR-Farbstoff wie SYBR® Green verwenden?

Bitte beachten Sie, dass SNP PolTaq DNA-Polymerase nicht für Real Time-PCR mit einem qPCR-Farbstoff geeignet ist. SNP Pol DNA-Polymerase funktioniert sehr gut mit qPCR-Farbstoffen.

Für welche Amplikonlänge sollten Primer ausgelegt werden?

Wir empfehlen, Primer mit einer kurzen Amplikonlänge (ca. 60-200 bp) zu entwerfen, um optimale Ergebnisse zu erzielen. Es sind jedoch auch längere Amplikonlängen möglich.