Introduction

The HiPure Plasmid DNA Mega Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 10 mg high copy number plasmid DNA can be purified from 500 mL overnight culture.

Details

Specifications

Features	Specifications
Main Functions	Isolation up to 10mg endotoxin-free plasmid DNA from 500ml bacterial culture
Applications	Cell transfection, animal injection, etc.
Purification method	Mega spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	High copy plasmid vector
Sample amount	500ml LB
Yield	1~10mg
Elution volume	≥2ml
Time per run	≤70 minutes
Liquid carrying volume per column	50ml
Binding yield of column	10mg

Principle

The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparation and clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
• Fast – silica gel column purification is much faster than ion exchange
• High yield – up to 10mg plasmid can be binded in one column
• Low endotoxin – the obtainedplasmid can be directly used for cell transfection and animal injection, etc.

Kit Contents

Contents	P111602	P111603
Purification Times	10 Preps	50 Preps
RNase A	60 mg	2 x 150 mg
Buffer E1	220 ml	2 x 550 ml
Buffer E2	220 ml	2 x 550 ml
Buffer E3	220 ml	2 x 550 ml
Buffer E4	220 ml	2 x 550 ml
Buffer E5	120 ml	550 ml
Buffer PW2	25 ml	2 x 100 ml
Elution Buffer	30 ml	120 ml
HiPure EF Maxi Columns	10	50
Lysate Clear Maxi Syringe	10	50
50 ml Collection Tubes	20	100

Storage and Stability

The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve.

Purchase Guide

Name	CAT NO	Features	Sampleamount	Elutionvolume	Yield
HiPure Plasmid Mini Kit	P1001	Classic, Regular application	1-5ml LB	30-50μl	5–35µg/5ml
HiPure Plasmid Mini Kit II	P1002	Classic, Regular application	5-15ml LB	75-100μl	20-80µg/15ml
HiPure Plasmid Plus 96 Kit	P1006	Classic, Regular application	1-1.5ml LB	70-150μl	1-15μg/1ml
HiPure Fastfilter Plasmid Midi Kit	P1013	Fast	30-50ml LB	0.3-0.8ml	50–250µg/50ml
HiPure Fastfilter Plasmid Maxi Kit	P1014	Fast	100-200ml LB	0.5-1.5ml	0.4–1mg/200ml
Low endotoxin/Endotoxin-free Plasmid
HiPure Plasmid EF Maxi Kit	P1156	Classic method, General for all kinds of vectors	100-200ml LB	≥0.7ml	200–1500µg
HiPure Plasmid EF Mini Kit	P1154	Recommend for low copy vector, Thoroughly remove RNA	5-15ml LB	≥60μl	10–80µg
HiPure Plasmid EF 96 Kit	P1157	Recommend for low copy vector, Thoroughly remove RNA	1-5ml LBx96	≥75μl	30µg
HiPure Plasmid EF Midi Kit	P1231	Recommend for High copy vector, Low elution volume, High concentration	25-50ml LB	≥100μl	10-500μg
HiPure Plasmid EF Mega Kit	P1116	Recommend for High copy vector(>3μg/ml)	500ml LB	≥2ml	1-10mg

For any technical problems or customized products, please contact us.

F&Q about Endotoxin-free Plasmid Extraction Kit — P1156 ←click here

The HiPure Plasmid DNA Mega Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 10 mg high copy number plasmid DNA can be purified from 500 mL overnight culture.

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	Plasmid Maxi preparation
Preservation conditions	Room temperature
Stability	Up to 4 years
Filter membrane	High quality glass fiber filter GF/B, 8 layers
Membrane aperture	1.0 μm
Maximum binding yield of plasmid	1 mg
Maximum yield of alcohol mediated binding	5 mg
Plasmid yield	Up to 1mg
Single liquid carrying capacity of column	20 ml
Minimum elution volume	800 μl
Withstand centrifugal force	3,000-5,000 x g
Centrifuge	Low speed centrifuge for 50ml centrifuge tubes, >3000 x g, swing-out Rotor

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
C13123	HiPure DNA Maxi Column III (8 x GF/B)with 50ml Collection Tubes	100/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.

Description

The SMOChem™ Deoxynucleotide (dNTP) Mix is an aqueous solution that contains an equimolar solution of ultrapure dATP, dCTP, dGTP and dTTP, each at a concentration of 10 mM at pH 8.5. The dNTP Mix is designed for many different molecular biology applications that involved in DNA synthesis or labeling, such as PCR, real-time PCR, DNA sequencing, reverse transcription, primer extension, and etc. The dNTP Mix is free of exo-deoxyribonuclease and endo-deoxyribonuclease as well as ribonuclease activity. The dNTP Mix offers the possibility to reduce the number of pipetting steps and the risk of reaction set up errors. 

Features

• Ideal for PCR amplification and cDNA synthesis
• Premixed solution
• Nuclease and ribonuclease free

Applications 

• PCR amplification of DNA fragments
• DNA fill-in reaction
• DNA sequencing
• Reverse transcription
• One-step RT-PCR

Storage

-20°C for 36 months

The SMOChem™ Deoxynucleotide (dNTP) Mix is an aqueous solution that contains an equimolar solution of ultrapure dATP, dCTP, dGTP and dTTP, each at a concentration of 10 mM at pH 8.5. The dNTP Mix is designed for many different molecular biology applications that involved in DNA synthesis or labeling, such as PCR, real-time PCR, DNA sequencing, reverse transcription, primer extension, and etc. The dNTP Mix is free of exo-deoxyribonuclease and endo-deoxyribonuclease as well as ribonuclease activity. The dNTP Mix offers the possibility to reduce the number of pipetting steps and the risk of reaction set up errors.