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P1116

CAT NO PRODUCT NAME SIZE PRICE
P111602 HiPure Plasmid EF Mega Kit 10 preps $298.00
P111603 HiPure Plasmid EF Mega Kit 50 preps $1,243.00
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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
Conventional plasmid, plasmid less than 30KB
Sample amount High copy plasmid: 500ml culture medium
Yield 10mg
Elution volume ≥0.8ml
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

Sample amount

Column type

Elution volume

Yield

Time per run

Centrifuge requirements

HiPure Plasmid EF Mini Kit

P1154 10-15ml 1.5ml column 75μl

10–70µg/15ml

≤40 minutes small centrifuge

HiPure Plasmid EF 96 Kit

P1157 1-5ml 96 well plate 75μl

30µg/1ml

≤60 minutes 96 well plate centrifuge

HiPure Plasmid EF Midi Kit

P1231 30-50ml 15ml column ≥0.4ml

10-500μg/50ml

≤60 minutes 15ml centrifuge

HiPure Plasmid EF Maxi Kit

P1156 100-200ml 50ml column ≥0.8ml

100–1500µg/200ml

60 minutes
50ml centrifuge

HiPure Plasmid EF Mega Kit

P1116
500ml
50ml column
≥0.8ml

10mg/500ml

70 minutes
50ml centrifuge
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