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HiDi DNA Polymerase

  • HiDi DNA polymerase is the gold-standard enzyme, whenever High Discrmination (hence the name "HiDi") is required. Benchmarking data with other well-known enzymes on the market show that HiDi is the best choice for highly selective PCRs, such as allele-specific PCR, HLA genotyping, analysis of single CpG methylation sites or the detection of mutations in a high background of wild-type sequences. 
    Whereas many DNA polymerases tolerate mismatched primer-template complexes, HiDi DNA polymerase efficiently discriminates those and produces specific amplicons in case of perfectly matched primer pairs. 

    The HiDi Taq variant has a 5'-3'-nuclease activity and therefore is suitable for hydrolysis probe-based assays.

  • HiDi and HiDi Taq DNA polymerase is hotstart-formulated with an aptamer. Temperatures above 50-55°C cause the aptamer’s secondary structure to melt and will set-free the polymerase. 
    HiDi is supplied as a 5 U/µl solution containing glycerol and is supplied together with 10x reaction buffer which has been optimized for short amplicons between 50-200 bp of length.
    The buffer contains 2.5 mM Magnesium-ions in the final 1x concentration. Additional Magnesium (+ 0.5 - 1.5 mM) might be added in case of longer amplicons >500 bp.
    HiDi DNA polymerase can also be used for real-time cycling, when adding a suitable real-time dye, for example GreenDye (#2000).

  • This product is shipped on cool packs. It is recommended to store the product upon arrival at -20°C.

  • HiDi DNA polymerase is tested for successful ASA performance detecting a genomic SNP (rs72921001) in HeLa genomic DNA. PCR products were subsequently analysed on a 2.5% agarose gel. A specific product was visualized by ethidium bromide staining at the correct amplicon length of 109 bp for the matched primer. In case of the mismatch primer no product formation was observed after 50 cycles.  

    HiDi Taq DNA polymerase was tested successfully for hydrolysis probe based real-time PCR. The product demonstrated linearity of amplification over a specified serial dilution of human genomic DNA. 

    The activity of HiDi and HiDi Taq DNA polymerase was monitored and adjusted to a specific DNA polymerase activity using an artificial DNA template and a DNA primer. Enzyme concentration was determined by protein-specific staining. Please inquire more information at info@mypols.de for the lot-specific concentration. No contamination was detected in standard test reactions.

Example Primer Design

Matching vs. mismatching nucleotide is placed at the 3'-end of the primer for best discrimination results.

Example Results - There´s no accounting for taste

Cilantro: some people love it in their food, some hate it. Here we are detecting a genomic SNP (rs72921001) in HeLa genomic DNA. This SNP is reported to be close to a number of genes coding for olfactory receptors. (Reference: Eriksson N. et al. (2012), "A genetic variant near olfactory receptor genes influences cilantro preference.")

Considering, that only the C-allele specific primer is extended and yielding in a specific amplicon, we can conclude a genetic predisposition in disliking cilantro, as this SNP is significantly associated with detecting a soapy taste to cilantro.  

Allele-specific PCRs were performed from 1 ng/µl of HeLa gDNA in the presence of a realtime dye, indicating the amplification of the C-allele specific primer only. The A-allele specific primer is discriminated, thus not amplified up to 50 cycles.

PCR products were subsequently analysed on a 2.5% agarose gel. Specific product is visualized by ethidium bromide staining at the amplicon length of 109 bp.

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