Introduction

The HiPure Fastfilter Plasmid DNA Midi 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. This system also includes aspecial filter cartridge which replaces the centrifugation step following alkaline lysis. 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 250 µg high copy number plasmid DNA or 10-75 µg low copy number plasmid DNA can be purified from 15-50 mL overnight culture.

Details

Specifications

Features	Specifications
Main Functions	Isolation up to 250μg plasmid DNA from 30-50ml bacterial culture
Applications	Enzyme digestion, sequencing, PCR and labeling,  etc.
Purification method	Midi spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Conventional plasmid, plasmid less than 30KB
Sample amount	30-50ml
Yield	50-250µg
Elution volume	≥300μl
Time per run	≤60 minutes
Liquid carrying volume per column	4ml
Binding yield of column	250µg

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: Preparationand 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 – it takes only 15-60 minutes to complete the isolation
• High yield – up to 70µg plasmid can be binded in one column

Kit Contents

Contents	P101302	P101303
Purification Times	25 Preps	100 Preps
RNase A	10 mg	40 mg
Buffer P1	80 ml	300 ml
Buffer P2	80 ml	300 ml
Buffer LEN3	40 ml	150 ml
Buffer GBT	60 ml	250 ml
Buffer PW1	60 ml	250 ml
Buffer PW2	20 ml	100 ml
Elution Buffer	30 ml	120 ml
HiPure DNA Midi Columns III	25	100
Lysate Clear Midi Syringe	25	100
15 ml Collection Tubes	50	200

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. After addition of RNase A，Buffer P1 is stable for 6 months when stored at 2-8°C.

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 Fastfilter Plasmid DNA Midi 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. This system also includes aspecial filter cartridge which replaces the centrifugation step following alkaline lysis. 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 250 µg high copy number plasmid DNA or 10-75 µg low copy number plasmid DNA can be purified from 15-50 mL overnight culture.

Description 

The DM5100 ExcelBand™ Super Range DNA Ladder is a ready-to-use DNA ladder, which is pre-mixed with loading dye for direct gel loading. The DNA Ladder DM5100 is composed of 26 individual DNA fragments: 25k, 10k, 8k, 6k, 5k, 4k, 3k, 2.5k, 2k, 1.5k, 1.2k, 1k, 900, 800, 700, 600, 500, 450, 400, 350, 300, 250, 200, 150, 100 and 50 base pairs derived from a mixture of PCR products and specifically digested plasmid DNA. This product contains four enhanced bands (3k, 1.2k, 500 and 200 bp) for easier reference. In addition, three tracking dyes, Xylene cyanol FF, Bromophenol blue and Orange G which mimic the migration of 4,000 bp, 500 bp and 50 bp dsDNA during electrophoresis are added for real time monitoring.

Features

• Sharp bands
• Quick reference— enhanced bands 
• Ready-to-use— premixed with loading dye for direct loading
• Stable— room temperature storage over 6 months

Source 

Phenol extracted PCR products and dsDNA digested with specific restriction enzymes, equilibrated in 10 mM Tris-HCl (pH 8.0) and 10 mM EDTA. 

Range 

50 ~ 25,000 bp 

Concentration 

88.7 µg/ 500 µl 

Recommended loading volume 

5 µl/ well 

Storage

Room temperature for 6 months
4°C for 12 months 
-20°C for 36 months