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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.
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.
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 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 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.
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.
- 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
high single nucleotide discrimination
SNP Pol DNA polymerase is supplied as a 5 U/µL solution. It comes together with an optimized 10x reaction buffer.
SNP Pol DNA polymerase shows no 5´-3´ exonuclease activity! Not applicable for usage together with hydrolysis probes for, e.g., TaqManTM probes.