Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Primer and probe mix (150 reactions)
Reverse Transcription, target specific primers (RNA genome viruses only)
Copy number standard curve (sufficient for multiple standard curves)
Internal extraction control – Read through VIC channel*
Endogenous control (150 tests)
RNAse/DNAse free water
*alternative fluorophores available on request

Overview

• Isolate genomic DNA from either exfoliated cells or bacteria found in urine
• Isolate and detect genomic DNA from as little as 1 mL of urine and up to 50 mL urine
• High quality DNA for sensitive applications
• Rapid processing time

Norgen’s Urine DNA Isolation Kit for Exfoliated Cells or Bacteria is designed for the rapid isolation of either: 1) human genomic DNA from exfoliated cells that have been shed into the urine from the urinary tract; or 2) bacterial genomic DNA from urine samples. The kit allows for the isolation of DNA from 1 to 50 mL of urine. The genomic DNA isolated from exfoliated cells can be used in a number of diagnostic and research applications including the diagnosis and monitoring of bladder, kidney, or other urinary-tract cancers. Bacterial genomic DNA from both human urine samples and urine samples from animals can be isolated with this kit in order to study the levels and types of bacteria that are present. The kit allows for the isolation of genomic DNA from both Gram negative and Gram positive bacteria, including E. coli, Proteus spp., Klebsiella spp., Enterobacter spp., Serratia spp., Pseudomonas spp, Clostridial ssp. and Leptospirosis spp.Chlamydia trachomatis and Neisseria gonorrhoeae.

Typical yields of human genomic DNA from exfoliated cells will vary depending on the cell density of the urine sample, which is affected by a number of factors including health, diet and sex of the individual donating the urine. Typical yields of bacterial genomic DNA will vary depending on the urine sample and the bacterial species, if any, present in the urine. Healthy humans generally have < 10, 000 CFU of bacteria per mL of urine, and this kit is sufficiently sensitive to isolate and detect DNA from even this minimal amount of bacteria. The genomic DNA purified with this kit is fully digestible with all restriction enzymes tested, and is completely compatible with downstream applications such as PCR, qPCR and Southern Blot analysis.

Details

Supporting Data

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Kit Specifications
Minimum Urine Input	1 mL
Maximum Urine Input	50 mL
Time to Complete 10 Purifications	10 minutes (Plus a 30 minute incubation – Bacteria) (Plus a 15 minute incubation – Exfoliated)

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 2 years after the date of shipment when stored at room temperature.

Component	Cat. 47050 (50 preps)
Resuspension Solution A	20 mL
Lysis Buffer B	40 mL
Wash Solution A	18 mL
Elution Buffer B	8 mL
Proteinase K in Storage Buffer	1.2 mL
Micro Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

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.