HiDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and therefor, is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) that is also termed allele-specific amplification (ASA).
HiDi® 2x PCR Master Mix - ready to use mix simplifies your PCR setup. Only target-specific primers and template need to be added as the mix contains all components for a successful and reliable PCR. This ensures reproducible results, significantly reduces set-up times and the risk of pipetting errors.
This PCR mix is also available with a full-length Taq DNA polymerase with a nuclease domain, featuring 100% compatibility with hydrolysis probes (TaqMan® probes etc.).
Benchmarking with products of competitors conducted by us and others show that the HiDi® DNA polymerase family is the first choice for highly selective PCRs, such as genotyping by allele-specific PCR, HLA genotyping, analysis of single CpG methylation sites or the detection of mutations in a high background of wild-type sequences. By using HiDi® DNA polymerase, less than 10 copies of a mutation can be detected in a background of >10.000 wild-type copies straight away without any other tedious assay optimization.
It has also been shown that HiDi® DNA polymerase family is highly suitable for quality control in CRISPR-Cas or TALEN-based applications.
Several independently conducted studies show that HiDi® DNA polymerase is ideally suited for use in asPCR in numerous research areas ranging from mutation detection to genome editing. (read more)
For research use and further manufacturing.
In case you are aiming to use our RUO products as components or for your development of e.g. an IVD medical device, please contact us.
What should I consider?
- This mix contains an engineered DNA polymerase in an optimized reaction buffer and ultrapure dNTPs. HiDi® 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® DNA polymerase can also be used for real-time cycling, when adding a suitable real-time dye, for example GreenDye (#2000). For hydrolysis probe based assays, please refer to our HiDi Taq DNA polymerase or our HiDi Taq 2x PCR master mix.
How do I have to store the mix?
- Keep at -20°C
- Minimize the number of freeze-thaw cycles by storing the product in aliquots. For a day-to-day use, we recommend keeping an aliquot at 4°C.
Download: HiDi® DNA Polymerase - Application note
Lifestyle-specific S-nitrosylation of protein cysteine thiols regulates Escherichia coli biofilm formation and resistance to oxidative stress.
"... After 3–4 cycles, single clones were isolated and screened by PCR using a high discrimination HiDi polymerase (myPols) and primers with the targeted original or mutated nucleotide at the 3′ end (see Table S5). ..."
Barraud N, Létoffé S, Beloin C, Vinh J, Chiappetta G, Ghigo JM.NPJ Biofilms Microbiomes. 2021 Apr 13;7(1):34.
Link to publication: https://doi.org/10.1038/s41522-021-00203-w
HiDi® 2x PCR Master Mix is tested for successful allele-specific PCR detecting a genomic SNP (rs72921001) in HeLa genomic DNA. In case of the mismatch primer no specific product formation was observed after 50 cycles.
The activity of HiDi® DNA polymerase is monitored and adjusted to a specific DNA polymerase activity using a synthetic DNA template and DNA primer. Enzyme concentrations are determined by protein-specific staining. Please inquire more information using our contact form for lot-specific concentrations. No contamination is detected in standard test reactions
Matching vs. mismatching nucleotide is placed at the 3'-end of the primer for best discrimination results.
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.