Introduction

Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.

The series of nucleic acid columns produced by Magen Biotech are based on carefully selected imported glass fiber membranes (GF/B, GF/D, GF/F). Columns production processes such as polypropylene injection molding materials, injection molding process, and downstream membrane packing and compression rings are strictly controlled. This is to ensure that the column has extremely high adsorption capacity and long-term stability. Compared with conventional products on the market, Magen’s columns are with varieties, and binding rate will not change when stored at room temperature for 4 years.

Details

Specifications

Features	Specifications
Recommended application	Small amounts of nucleic acid isolation, viral nucleicacid from cell free samples
Preservation conditions	Room temperature
Stability	Up to 4 years
Filter membrane	High quality glass fiber filter GF/F, 3 layers
Membrane aperture	0.7μm
Maximum binding yield of plasmid	30 μg
Maximum yield of alcohol mediated Binding	200 μg
Single liquid carrying capacity of column	800 μl
Minimum elution volume	30 μl
Withstand centrifugal force	16,000 x g
Centrifuge	Small high speed centrifuge (2ml)

Adsorption Mechanism

Based on the negatively charged DNA skeleton, it has a high affinity for positively charged glass fibers. In high salt and ethanol solutions, DNA/RNA binds to glass fiber and interacts with hydrophilic matrix on silica through hydrogen bond. DNA/RNA is tightly bound. All pollutants can be removed by washing solution. At high salt concentration, nucleic acids selectively bind to silicagel membrane, while other pollutants, mainly proteins, are removed by membrane washing.

Ordering information

CAT.No.	Product Name	Package
C13112	HiPure Viral Mini Column I (3 x GF/F)with 2ml Collection Tubes	1000/Bag

Purchase Guide

Item No.	Product Name	Membrane type/number of layers	Collection tubes	Plasmid DNA binding capacity (Physical adsorption)	gDNA/RNA binding capacity (Alcohol-mediated adsorption)	Minimum Elution volume	Liquid volume capacity
C13010	HiPure DNA Nano Column	2 layers GF/F	2ml without cap	5μg	20μg	10μl	700μl
C13011	HiPure DNA Micro Column	3 layers GF/F	2ml without cap	10μg	50μg	15μl	700μl
C13100	HiPure DNA Mini Column I	2 layers GF/B	2ml without cap	15μg	100μg	30μl	700μl
C13110	HiPure DNA Mini Column II	4 layers GF/B	2ml without cap	35μg	200μg	50μl	800μl
C13111	HiPure RNA Mini Column	3 layers GF/B	2ml without cap	30μg	200μg	30μl	800μl
C13112	HiPure Viral Mini Column	3 layers GF/F	2ml without cap	30μg	200μg	30μl	800μl
C13113	HiPure CFDNA Mini Column	3 layers GF/F,1 layer GF/B	2ml without cap	30μg	200μg	30μl	800μl
C13120	HiPure DNA Midi Column	4 layers GF/B	15ml Centrifuge tube	125μg	1mg	500μl	4ml
C13121	HiPure DNA Midi Column III	8 layers GF/B	15ml Centrifuge tube	250μg	1mg	500μl	4ml
C13122	HiPure DNA Maxi Column	4 layers GF/B	50ml Centrifuge tube	500μg	5mg	1000μl	20ml
C13123	HiPure DNA Maxi Column III	8 layers GF/B	50ml Centrifuge tube	1mg	5mg	1000μl	20ml
C13124	HiPure DNA Maxi Column C	8 layers GF/B	50ml high speed Centrifuge tube	1mg	5mg	700μl	12ml
C13130	HiPure DNA Plate	2 layers GF/F	1.6ml Plate	30μg	100μg	80μl	900μl
C13131	HiPure gDNA Plate	2 layers GF/B	1.6ml Plate	30μg	100μg	80μl	900μl

Note: GF/B pore size is for 1.0μM glass fiber membrane; GF/F pore size is for 0.7μm glass fiber membrane.

Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.

• 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.

Introduction

This kit is suitable for high-throughput extraction of total RNA from paraffin embedded tissue samples, including miRNA. The kit is based on superparamagnetic magnetic particle purification technology. The extraction process does not require the use of toxic phenolic chloroform extraction or time-consuming alcohol precipitation.The whole extraction process only takes 120 minutes. The obtained RNA can be directly used in RT-PCR. The high-throughput extraction can be realized by 96 hole magnetic frame and 96 holeo scillator, various nucleic acid extractors (such as KingFisherFle, MagMix32, MagMix 96) or cooperating with the pipette workstation (Hamilton, Tican) for automatic extraction.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from FFPE tissue
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Products	RNA, miRNA
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Adaptive instrument	Nucleic acid extractor, pipetting workstation
Sample type	FFPE slice, FFPE embedded tissue
Sample amount	≤6 slices

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Protease. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• Fast – several samples can be extracted in 120 minutes by column method
• Safe – no phenol chloroform extraction required

Kit Contents

Contents	IVD3022
Purification Times	200 Preps
MagBind Particles	4.5 ml
Proteinase K	100 mg
Protease Dissolve Buffer	6 ml
DNase I	4 x 600 µl
DNase Buffer	30 ml
Buffer DPS	200 ml
Buffer FRL	40 ml
Buffer AL	40 ml
Buffer MW1*	110 ml
Buffer MW2*	2 x 50 ml
Nuclease Free Water	30 ml

Storage and Stability

Proteinase K and MagBind Particles should be stored at 2–8°C upon arrival. DNase I should bestored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K and MagBind Particles up to 8 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under theseconditions.

This kit is suitable for high-throughput extraction of total RNA from paraffin embedded tissue samples, including miRNA. The kit is based on superparamagnetic magnetic particle purification technology. The extraction process does not require the use of toxic phenolic chloroform extraction or time-consuming alcohol precipitation.The whole extraction process only takes 120 minutes. The obtained RNA can be directly used in RT-PCR. The high-throughput extraction can be realized by 96 hole magnetic frame and 96 holeo scillator, various nucleic acid extractors (such as KingFisherFle, MagMix32, MagMix 96) or cooperating with the pipette workstation (Hamilton, Tican) for automatic extraction.