Introduction

A highly efficient, easily automated PCR purification system that delivers superior quality DNA with no salt carryover. Requiring no centrifugation or filtration. This kit can be easily used in manual and automated 96 or 384-well formats.

Details

Specifications

Features	Specifications
Main Functions	Recover 100-400μl DNA/ RNA from PCR products / enzymatic reaction solution / or crude DNA / RNA
Applications	Automatic sequencing, enzyme digestion, PCR and labeling
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Agarose Gel, PCR products, enzymatic reaction solution
Sample amount	100-400µl
Recovery	80%
Elution volume	≥30μl

Principle

The MagPure method contains magnetic particles in an optimized binding buffer to selectively bind DNA fragments 100bp and larger toparamagnetic beads. Excess primers, nucleotides, salts, and enzymes can be removed using a simple washing procedure.

Advantages

• High recovery efficiency – up to 80% DNA recovery

Kit Contents

Contents	D500301	D500302	D500303
Purification Times	50 Preps	500 Preps	5000 Preps
Buffer AL	10 ml	60 ml	550 ml
Buffer BD*	5 ml	25 ml	2 x 100 ml
MagPure RNA Particles	1.2 ml	12 ml	120 ml
RNase Free Water	5 ml	30 ml	250 ml

Storage and Stability

MagPure RNA Particles should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

A highly efficient, easily automated PCR purification system that delivers superior quality DNA with no salt carryover. Requiring no centrifugation or filtration. This kit can be easily used in manual and automated 96 or 384-well formats.

Overview

• Clean-up & concentrate total RNA (including miRNA) in minutes
• Clean-up & concentrate RNA from TRIzol^®, TRI Reagent^®, etc
• Clean-up RNA from contaminants including enzymes, primers, nucleotides
• Rapid spin-column protocol, elute in 20 µL
• 96-well format for high throughput processing & Micro-Elute spin column formats for 8 µL are available
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits are designed to clean-up and concentrate RNA (including miRNA) from TRIzol^® and TRI Reagent^®, enzymatic reactions, in vitro transcription, labelling reactions, etc. These are robust kits for all clean-up and concentration purposes for up to 35 µg of RNA in solution. The purified RNA is of the highest purity and integrity, and can be used in a number of downstream applications. These kits purify all sizes of RNA, from large mRNA and ribosomal RNA down to miRNA, siRNA and lncRNA.

RNA Clean-Up and Concentration Kit (Spin Column)

Norgen’s RNA Clean-Up and Concentration Kit provides a rapid method for the purification, cleanup and concentration of up to 35 μg of RNA isolated using different methods including phenol/guanidine-based protocols, and from various upstream enzymatic reactions such as DNase treatment and labeling. The kit elutes RNA in 20 µL. Complete 10 purifications in 20 minutes.

RNA Clean-Up and Concentration 96-Well Kit (High Throughput)

Norgen’s RNA Clean-Up and Concentration 96-Well Kit provides a rapid method for the purification, cleanup and concentration of up to 50 μg of RNA isolated using different methods including phenol/guanidine-based protocols, and from various upstream enzymatic reactions such as DNase treatment and labeling. The kit elutes RNA in 75 µL. Complete 10 purifications in 30 minutes.

RNA Clean-Up and Concentration Micro-Elute Kit (Micro-Elute)

Norgen’s RNA Clean-Up and Concentration Micro-Elution Kit provides a rapid method for the purification, cleanup and concentration of up to 10 μg of RNA isolated using different methods including phenol/guanidine-based protocols, and from various upstream enzymatic reactions such as DNase treatment, labeling and in vitro transcription. This kit is made for eluting RNA in smaller volumes of 8 µL for all types of downstream applications, for the highest concentration of RNA sample. Complete 10 purifications in 20 minutes.

Protocol

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	10 μg
Size of RNA Purified	All sizes, including small RNA (<200 nt)
Maximum Amount of Starting Material	10 μg of RNA
Minimum Elution Volume	8 μL
Time to Complete 10 Purifications	20 minutes
Average Recovery	≥ 90%

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.

Component	Cat. 23600 (50 preps)	Cat. 43200 (100 preps)	Cat. 25100 (192 preps)	Cat. 61000 (50 preps)
Buffer RL	40 mL	2 x 40 mL	2 x 40 mL	40 mL
Wash Solution A	38 mL	2 x 38 mL	2 x 38 mL	38 mL
Elution Solution A	6 mL	2 x 6 mL	2 x 20 mL	6 mL
Column Activation Solution	–	–	–	30 mL
Micro Spin Columns	50	100	–	–
Micro-Elute RNA Spin Columns	–	–	–	50
96-Well Plate	–	–	2	–
Adhesive Tape	–	–	4	–
Collection Tubes	50	100	–	50
96-Well Collection Plate	–	–	2	–
Elution Tubes (1.7 mL)	50	100	–	50
96-Well Elution Plate	–	–	2	–
Product Insert	1	1	1	1

• 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.