Introduction

The HiPure Midi system provides a fast, simple, and cost-effective plasmid DNA midiprep method for routine molecular biology laboratory applications. HiPure Midiprep Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.

Details

Specifications

Features	Specifications
Main Functions	Isolation up to 500µg endotoxin-free plasmid DNA from 25-50ml bacterial culture. Recommend for High copy vector, Low elution volume, High concentration
Applications	Cell transfection, animal injection, etc.
Purification method	Midi spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	High copy plasmid vector
Sample amount	25-50ml LB
Yield	10-500μg
Elution volume	≥0.1ml
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: 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 concentration – easily obtain concentrations >1000ng/μl
• Small amount large extraction – using a small amount of column to achieve a large extraction yield (500μg)
• Low endotoxin content – the obtained plasmid can be directly used in cell transfection and animal injection

Kit Contents

Contents	P123102	P123102B
Purification Times	50 Preps	50 Preps
RNase A	30 mg	30 mg
Buffer E1	150 ml	150 ml
Buffer E2	150 ml	150 ml
Buffer E3	150 ml	150 ml
Buffer E4	150 ml	150 ml
Buffer E5	150 ml	150 ml
Buffer PW2*	50 ml	50 ml
Elution Buffer	30 ml	30 ml
MaxPure EF Mini Column	50	50
2 ml Collection Tubes	50	50
Lysate Clear Midi Syringe	50	50
Extend Tubes	50	50
50ml Centrifuge Tubes		50
Support Tubes		50

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 and optional LyseBlue reagent, 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 Midi system provides a fast, simple, and cost-effective plasmid DNA midiprep method for routine molecular biology laboratory applications. HiPure Midiprep Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.

Overview

• Rapid isolation of both small and large species of DNA from urine
• Convenient spin column format
• Effective removal of PCR inhibitors
• Purified DNA is highly suited to sensitive downstream applications
• Allows for the purification of viral DNA from urine

Both high molecular weight DNA (greater than 1 kb in size; mostly cell associated) and the smaller DNA (150 – 250 bp; derived from the circulation) is effectively isolated and purified using a rapid and convenient spin column protocol. This kit can be used to isolate DNA from a broad range of viruses in urine as well. Salts, metabolic wastes, proteins and other contaminants are removed to yield inhibitor-free DNA for use in sensitive applications. The DNA is of excellent quality for various downstream applications such as PCR, qPCR and DNA fingerprinting, methylation studies and more.

This kit is fully compatible with Norgen’s Urine Collection and Preservation Tubes.

Urine DNA Isolation Kit

This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 50 μL to 1.75 mL of urine. Multiple samples can be processed in 30 minutes.

Urine DNA Isolation Kit (Slurry Format)

This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 3 mL to 25 mL. Multiple samples can be processed in 30 minutes. Multiple samples can be processed in 45 minutes.

Urine DNA Isolation Maxi Kit (Slurry Format)

This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 25 mL of urine up to 80 mL. Multiple samples can be processed in 45 minutes.

Background

DNA found in urine can be divided into 2 basic categories. The larger species, genomic-DNA (gDNA), is generally greater than 1 kb in size, and appears to be derived mainly from exfoliated cells. The second species is smaller, generally between 150 and 250 bp (apoptotic-DNA), and derives, at least in part, from the circulation. The second species is also considered as an RNA/DNA hybrid as reported by Halicka et al. (2000). Both types of DNA can be isolated reliably using this kit.

Details

Supporting Data

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Kit Specifications – Spin Column
Minimum Urine Input	3 mL
Maximum Urine Input	25 mL
Time to Complete 10 Purifications	< 30 minutes
Size of Urine DNA Purified	Large (> 1 kb) and small (150-250 bp)

Kit Specifications – 96-well
Urine Input Volume	3 – 25 mL
Elution Volume	1st Elution 100 μL 2nd Elution 100 μL
Size of DNA Purified	Large (> 1 kb) and small (150-250 bp)
Time to Complete 96 Purifications	< 1 hour

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm up Slurry B1 and Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed. Slurry B1 contains a grey resin that will not dissolve when warmed.

Component	Cat. 18100 (50 preps)	Cat. 48800 (50 preps)	Cat. 50100 (50 preps)
Binding Solution K	15 mL	–	–
Slurry B1	–	18 mL	18 mL
Proteinase K in Storage Buffer	2 mL	–	–
Pronase K in Storage Buffer	2 mL	–	–
Soluton WN	9 mL	–	–
Binding Buffer A	–	–	50 mL
Lysis Buffer A	–	30 mL	30 mL
Wash Solution A	–	38 mL	38 mL
Wash Solution B	30 mL	–	–
Wash Solution D	9 mL	–	–
Binding Solution K	15 mL	–	–
Elution Buffer B	15 mL	15 mL	15 mL
Micro Spin Columns	50	–	–
Mini Filter Spin Columns	–	50	50
Collection Tubes	50	50	50
Elution Tubes (1.7 mL)	100	100	100
Product Insert	1	1	1