IST-300 96 Extraction Tube Sealing Film

• 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

• Sequentially purify total RNA (and miRNA), DNA and proteins from a single sample
• No sample splitting or need to use phenol or precipitation methods
• Purify RNA/DNA/Protein from cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

RNA/DNA/Protein Purification Plus Kit

This kit provides a rapid method for the high throughput isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, or yeast. The kit employs two columns: 1) for gDNA purification and 2) for RNA purification utilizing Norgen’s resin columns (superior for the binding of all RNA sizes including miRNA). The proteins are also purified on the second column after RNA elution. The proteins are eluted in buffer and are ready for downstream application without any further clean up required. The proteins can be quantified directly, used in western blots, ELISA or mass spectrometry. This kit provides a rapid spin-column method for the isolation and purification of total RNA, genomic DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants.

RNA/DNA/Protein Purification Plus Micro Kit

This kit provides a rapid spin-column method for the isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, microdissected samples including LCM, stem cells, sorted cells, and CTC. The total RNA, genomic DNA and proteins are all column purified in less than 30 minutes. The RNA and DNA can be eluted in as little as 20 µL while the protein can be eluted in as little as 50 µL. This kit provides the same performance as if the samples were isolated from dedicated kits.

RNA/DNA/Protein Purification 96-Well Plus Kit

The kit employs two plates: 1) for DNA purification and 2) for RNA purification utilizing Norgen’s resin (superior for the binding of all RNA sizes including miRNA). Please see the protocol schematic below.

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	35 μg for RNA 10 μg for DNA 100 μg for protein
Maximum Column Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Size of DNA Purified	≥ 30 kb
Time to Complete 10 Purifications	30 minutes
Maximum Amount of Starting Material:Animal CellsAnimal TissuesBloodPlant / Fungi Tissues	5 x 105 cells5 mg (for selected tissues)50 µL25 mg
Average Yield* HeLa Cells (5 x 105 cells) HeLa Cells (5 x 105 cells) HeLa Cells (5 x 105 cells)	RNA: 7.5 μg DNA: 1-2 μg Protein: 70 μg

Storage Conditions and Product Stability
The Protein Loading Dye should be stored at -20°C after the addition of DL-Dithiothreitol (DTT). 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. 47700 (50 preps)	Cat. 51600 (50 preps)	Cat. 51700 (96 preps)
Buffer SKP	40 mL	40 mL	–
Lysis Buffer Q	–	–	40 mL
Wash Solution A	2 x 38 mL 1 x 18 mL	2 x 38 mL 1 x 18 mL	2 x 38 mL 1 x 18 mL
Elution Solution A	6 mL	6 mL	20 mL
Elution Buffer F	15 mL	15 mL	2 x 15 mL
Wash Solution C	30 mL	30 mL	60 mL
Binding Buffer A	8 mL	8 mL	8 mL
Elution Buffer C	8 mL	8 mL	30 mL
Protein Neutralizer	4 mL	4 mL	4 mL
Protein Loading Dye	2 mL	2 mL	3 x 2 mL
gDNA Purification Columns	50	–	–
gDNA Purification Micro Columns	–	50	–
gDNA Purification 96-Well Plate	–	–	1
RNA/Protein Purification Columns	50	–	–
RNA/Protein Purification Micro Columns	–	50	–
RNA/Protein Purification 96-Well Plate	–	–	1
Collection Tubes	150	150	–
Collection Plate	–	–	5
Elution Tubes (1.7 mL)	150	150	–
Elution Plate	–	–	3
Lysis Preparation Plate	–	–	2
Adhesive Tape	–	–	4
Product Insert	1	1	1

1.5 mL – 2 mL Micro-centrifuge Tube or Screw-Cap Vial Chiller Block

SKU: CB024

Permagen’s SBS/ANSI footprint 24-Tube thermal block allows the use of up to twenty four 1.5 mL -2 mL Microcentrifuge tubes, or screw-cap vials for keeping sample temperatures consistent. Can be used for heating as well

Permagen’s SBS/ANSI footprint 24-Tube thermal block allows the use of up to twenty four 1.5 mL -2 mL Microcentrifuge tubes, or screw-cap vials for keeping sample temperatures consistent. Can be used for heating as well