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HiDi DNA Polymerase

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Manufacturer: Genaxxon bioscience



Order number: M3009.0000

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Product information "HiDi DNA Polymerase"

For allele specific discrimination, eg. of CRISPR/Cas9 point mutations or for the identification of wrong CRISPR/Cas9 products or for verification of sequencing results. HiDi and HiDiTaq DNA polymerase efficiently discriminate primers, which have a mismatch at the 3'-end! HiDiTaq 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
HiDi 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 HiDi DNA polymerase, on the other hand, distinguishes these specifically (high discrimination) and supplies only PCR products with perfectly matching primer pairs! The Genaxxon HiDi and HiDiTaq 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 HiDiTaq or HiDi 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 HiDi DNA polymerase. The HiDiTaq DNA polymerase is very suitable for the analysis of liquid biopsy samples. With HiDiTaq, the presence and frequency of cancer mutations can be analyzed and quantified very well. 

Picture below: Application note HiDi 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 HiDi 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 HidiTaq 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) we can offer additional products for your PCR.

Application areas for HiDi DNA and HiDiTaq 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
- Multiplex PCRs
- 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

 

> frequently asked questions (FAQs)

> Online form for your HiDi test sample

 

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 - Multiplex PCRs - realtime PCR with hydrolysis probes - realtime multiplex PCRs
 
 

Our comment on "HiDi DNA Polymerase"

high single nucleotide discrimination
 

Technical Data:

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

HiDi DNA polymerase shows no 5’-3’ exonuclease activity! Not applicable for usage 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 - Multiplex PCRs - 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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