Description

Escherichia coli is one of many species of bacteria living in the lower intestines of mammals, known as gut flora. When located in the large intestine, it assists with waste processing, vitamin K production, and food absorption. Discovered in 1885 by Theodor Escherich, a German pediatrician and bacteriologist, E. coli are abundant: the number of individual E. coli bacteria in the faeces that a human defecates in one day averages between 100 billion and 10 trillion. However, the bacteria are not confined to the environment, and specimens have also been located, for example, on the edge of hot springs. The E. coli strain O157:H7 is one of hundreds of strains of the bacterium that causes illness in humans.

E. coli are unable to sporulate. Thus, treatments which kill all active bacteria, such as pasteurization or simple boiling, are effective for their eradication, without requiring the more rigorous sterilization which also deactivates spores. As a result of their adaptation to mammalian intestines, E. coli grow best in vivo or at the higher temperatures characteristic of such an environment, rather than the cooler temperatures found in soil and other environments.

The enteric E. coli (EC) are divided on the basis of virulence properties into enterotoxigenic (ETEC – causative agent of diarrhea in humans, pigs, sheep, goats, cattle, dogs, and horses), enteropathogenic (EPEC – causative agent of diarrhea in humans, rabbits, dogs, cats and horses); enteroinvasive (EIEC – found only in humans), verotoxigenic (VTEC – found in pigs, cattle, dogs and cats); enterohaemorrhagic (EHEC – found in humans, cattle, and goats, attacking porcine strains that colonize the gut in a manner similar to human EPEC strains) and enteroaggregative E. coli (EAggEC – found only in humans).

E. coli O157:H7 was first recognized as a pathogen as a result of an outbreak of unusual gastrointestinal illness in 1982. The outbreak was traced to contaminated hamburgers, and the illness was similar to other incidents in the United States and Japan. The etiologic agent of the illness was identified as a rare O157:H7 serotype of Escherichia coli in 1983. This serotype had only been isolated once before, from a sick patient in 1975.

Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target Accurate controls to confirm findings

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

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

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

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.

• ASTM Type I Water
• Produced by reverse osmosis, ion-exchange, electro-deionization, ultrafiltration, UV treatment (UVC & 185 nm), 0.1 μm membrane filtration
• Nuclease-free, protease-free, endotoxin-free
• Suitable for molecular biology applications (qPCR, NGS etc.)
• Suitable for cell culture applications
• Produced in an ISO certified GMP laboratory
• Water system is routinely monitored for compliance with QC standards

The Norgen water purification system has been designed in conjunction with experts from Millipore Sigma. The Ultrapure NFW is purified in a two-stage process. The first stage, to produce type II water, includes pre-treatment with activated charcoal and polyphosphate, this water then undergoes reverse osmosis, electro-deionization and ion exchange to remove contaminants as well as UV treatment for disinfection. The Pure water then flows to a second system in a clean room with an additional 4 layers of purification. After passing through an additional ion exchange column, the water is then subjected to Ultrafiltration to remove particulates, bacteria, & ionic contaminants down to trace level, the water is then treated with UV light (182 nm wavelength) to break down organic matter. Lastly a second Ultrafilter for the production of pyrogen-, nuclease- and bacteria-free water at 18.2 MΩ. This water is of utmost purity and suitable for direct use in molecular biology applications such as qPCR and NGS, as well as cell culture and other applications.

Details

Applications
For use in any molecular biology application

Storage Conditions
Norgen’s Nuclease-Free Water should be stored at room temperature. Storage at +4°C or -20°C is optional.

Precautions and Disclaimers
This product is designed for research purposes only.  It is not intended for human or diagnostic use.