Checkit Go Volumetric Verification Kit Products

dsDNA Quantification BR Kit

The kit includes BR Dye, BR Dilution Buffer, and two DNA Standards. The assay is accurate for DNA concentrations from 100 pg/µL to 1000 ng/µL, and is highly selective for double-stranded DNA over RNA.

Features

• • • Kit was optimized for use with the Qubit® Fluorometer
    • Uses the Qubit® dsDNA Broad Range assay setting
    • Linear range is 2-1000 ng dsDNA
    • Great saving since the kit cuts the costs by 60%

DNA selectivity and sensitivity. A series of input DNA (200, 400, 600, 800, and 1000 ng) was used with or without RNA contamination.

Cat.# 40041 dsDNA Quantification BR Kit, 100 assays and 500 assays

• 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

• Purify sequencing extension products from dye terminators, primers and other contaminants
• Also purify DNA from different enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations.
• High recovery
• Fast and efficient spin column format
• Also available in 96 well format for high throughput

This kit provides a rapid spin column procedure for the purification and clean-up of sequencing and various other enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations. The kit is used to remove reaction contaminants including dye terminators, salts, enzymes, excess primers and primer dimers. Contaminants are undesirable as they can interfere with many downstream applications including sequencing, RFLP, restriction enzyme digestions and ligation. Purification is based on spin-column chromatography without the use of phenol, chloroform or alcohol precipitation. The kit provides a high quality product with up to 90% recovery.

The kit is also available in a 96-well format for high-throughput sequencing reaction clean-up.  Purification with the 96-well plate can be performed using either a vacuum manifold or centrifugation.

Details

Supporting Data

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Kit Specifications – 96-well
Binding Capacity Per Well	15 μg
Maximum Loading Volume Per Well	500 μL
Average DNA Recovery	Up to 90%
Time to Complete 96 Purifications	20 minutes
Minimum Elution Volume	50 μL

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.

Component	Cat. 34500 (50 preps)	Cat. 34400 (2 x 96-well plate)
Binding Buffer C	30 mL	3 x 30 mL
Wash Solution A	12 mL	2 x 20 mL
Elution Buffer B	8 mL	2 x 15 mL
Spin Columns	50	–
Collection Tubes	50	–
Elution Tubes (1.7 mL)	50	–
96-Well Plate	–	2
Adhesive Tape	–	4
96-Well Collection Plate	–	2
96-Well Elution Plate	–	2
Product Insert	1	1