N-(Propargyl-PEG2)-DBCO-PEG3-Amine, TFA salt

• 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.

HiDi is available as:

HiDi® DNA Polymerase (>>)
HiDi® Taq DNA Polymerase (>>)
HiDi® 2x PCR Master Mix (>>)
HiDi® Taq 2x PCR Master Mix (>>)

Casestudies:
HiDi® DNA Polymerase: Applications from mutation detection to genome editing (read more)

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.

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.

Overview

• Able to quantify NGS Library (Illumina) of a wide spectrum of concentrations, including sub-nanomolar concentrations
• DNA is accurately quantified by using a standard curve constructed from the provided DNA Standards
• Specially designed DNA standards for Small RNA-Seq library; also compatible to NGS library of other molecular weights

Norgen’s NGS Library Quantification Kit (for Small RNA-Seq) offers a PCR-based detection procedure to quantify NGS libraries (specifically Small RNA-Seq) of a wide spectrum of concentrations.  The kit consists of a specially designed primer mix compatible with the Illumina system, that is used in conjunction with the provided 2X Real-Time PCR Master Mix to amplify a library of unknown concentration. The unknown library is accurately quantified by using a standard curve constructed from the provided DNA Standard (range from 20 pM to 2 fM) on a Real-Time PCR System. The kit is specially optimized to quantify Small RNA-Seq libraries with DNA standards that have similar size to a Small RNA-Seq library. However, it could also be used with other types of NGS libraries.

Details

Supporting Data

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Storage Conditions
Upon receipt, store Norgen’s NGS Library Quantification Kit (for Small RNA-Seq) at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.

Component	Cat. 61600 (100 reactions)
2X Real-Time PCR Master Mix	1 mL
NGS Library Quantification Primer Set Mix	600 µL
Quantified NGS Library Standards (for Small-RNA Seq)*	5 x 80 µL Standards
Product Insert	1

* Quantified Library Standards are provided as 20 pM, 2 pM, 200 fM, 20 fM and 2 fM

Real-time fluorescent DNA amplification made possible with a TwistAmp® exo kit. Recommended for users who want to combine TwistDx’s RPA amplification technology with the use of TwistDx’s proprietary fluorescent TwistAmp® exo probe in a homogenous format. The user need only supply primers, probe, and template. See manual for more information. Click to order oligonucleotides.

Real-time fluorescent DNA amplification made possible with a TwistAmp® exo kit. Recommended for users who want to combine TwistDx’s RPA amplification technology with the use of TwistDx’s proprietary fluorescent TwistAmp® exo probe in a homogenous format. The user need only supply primers, probe, and template. See manual for more information. Click to order oligonucleotides.