• 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 is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) or allele-specific amplification (ASA).Please note: This DNA polymerase is also available as a nuclease deficient variant, featuring higher robustness towards potential PCR inhibitors and compatibility with real-time dyes such as our GreenDye.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® Taq 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.HiDi® Taq DNA polymerase harbours a nuclease function and therefor is also suitable for use with hydrolysis probes (TaqMan® probes etc.). It has also been shown that HiDi® DNA polymerase family is highly suitable for quality control and mutation identification in CRISPR-Cas or TALEN-based applications.Several independently conducted studies show that HiDi® Taq 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 is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) or allele-specific amplification (ASA).

Please note: This DNA polymerase is also available as a nuclease deficient variant, featuring higher robustness towards potential PCR inhibitors and compatibility with real-time dyes such as our GreenDye.

Overview

• Isolate genomic DNA from all types of bacteria (both Gram-positive and Gram-negative)
• Rapid and convenient spin column protocol
• 96-well format available for high throughput
• High yield, high quality DNA for sensitive downstream applications including sequencing, PCR, qPCR and more

This kit is designed for the rapid spin column preparation of genomic DNA from 2 x 10⁹ viable bacterial cells (between 0.5 and 1.0 mL of culture). This kit can be used for both Gram-negative and Gram-positive bacteria including Escherichia coli and Bacillus cereus. Purified genomic DNA is of an excellent quality and yield, and is fully compatible with restriction enzyme digestions, sequencing, PCR, qPCR and more. Also available in 96-well format for high throughput applications.

Details

Supporting Data

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Kit Specifications (Spin Columns)
Input	2 x 109 bacterial cells
Column Binding Capacity	25 µg
Average Yield*	Up to 20 µg
Time to Complete 10 Purifications	1 hour

* Yield will vary depending on the type of sample processed

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 2 years after the date of shipment when stored at room temperature.

Component	Cat. 17900 (50 preps)	Cat. 17950 (192 preps)
Resuspension Solution A	20 mL	60 mL
Lysis Buffer P	18 mL	60 mL
Solution BX	28 mL	110 mL
Wash Solution A	18 mL	2 x 38 mL
Elution Buffer B	30 mL	2 x 30 mL
Proteinase K	12 mg	50 mg
Spin Columns	50	–
96-Well Plate	–	2
Adhesive Tape	–	4
Collection Plate	–	2
Collection Tubes	50	–
Elution Tubes (1.7 mL)	50	–
Elution Plate	–	2
Product Insert	1	1

DNA Concentrator (Magnetic Beads)

The DNA Concentrator (Magnetic Beads) can be used to efficiently concentrate DNA/RNA from various samples with low concentration without the need of a DNA vacuum concentrator. Solid Phase Reversible Immobilization (SPRI) magnetic beads are well used for DNA and RNA purification and concentration. The beads are paramagnetic particles coated with carboxyl groups that reversibly bind to DNA and RNA.

The concentrator protocol is simple: mix Concentrator Buffer and Concentrator Beads with the sample, wash, and elute pure DNA in a small volume to concentrate the DNA samples. Moreover, the DNA/RNA samples are also purified during the procedure. The beads with our unique technology purify DNA/RNA samples effectively by removing unwanted components such as dNTPs, enzymes, detergents, proteins, and other contaminants.

However, traditional magnetic beads can only bind nucleic acids that are 100 bp or longer. Nucleic acids shorter than 100 bp are not effectively recovered. We have successfully developed the kit that overcomes the hurdle of short nucleic acids recovery. The reagent can be used for concentration of samples from genomic DNA to short DNA/RNA.

Recovery rate

• >90% for DNA size >30 bp
• >80% for DNA size between 20-29 bp

Features

• Effective concentrator of DNA and RNA samples, even small DNA, RNA, oligos
• Binding capacity: 10 ug DNA per prep
• Removal of unwanted components and other impurities

The DNA Concentrator (Magnetic Beads) can be used to efficiently concentrate DNA/RNA from various samples with low concentration without the need of a DNA vacuum concentrator. Solid Phase Reversible Immobilization (SPRI) magnetic beads are well used for DNA and RNA purification and concentration. The beads are paramagnetic particles coated with carboxyl groups that reversibly bind to DNA and RNA.