Overview

• Quantified standard to be used as a positive control or PCR quantification standard
• Vigorously quantified using multiple methods
• Includes partial sequence of STX-1 subunit A, and STX-2 subunit A genes (please refer to product insert for more details)
• Compatible with E. coli O157:H7 TaqMan PCR Detection Kit and E. coli O157:H7 TaqMan PCR Detection Probe/Primer and Control Set

E. coli O157:H7 is a rod-shaped, gram negative bacterium. It is an enterohemorrhagic strain of the common E. coli bacterium and infection by the O157:H7 strain is commonly associated with hemorrhagic colitis. E. coli O157:H7 is recognized by its somatic (cell wall) antigen (O157) and its flagella antigen (H7). In addition, E. coli O157:H7 is known to produce Shiga-like toxins, which cause severe symptoms. While most patients can recover from the infection, up to 15% of the patients may develop hemolytic uremic syndrome, a type of kidney failure that could be fatal. Infection of E. coli O157:H7 usually results from consumption of poorly prepared food including undercooked meat (particularly ground beef), untreated water or raw unpasteurized milk.

Norgen’s E. coli O157:H7 Quantified Bacterial DNA Standards are cloned fragments of the two Shiga-like toxin regions purified using Norgen’s sample preparation technology. They include partial sequences of STX-1 subunit A gene and STX-2 subunit A gene. Please refer to Appendix A for sequence information. The purified DNA is quantified vigorously using multiple methods including spectrophotometry, gel densitometry and real-time PCR. It is intended to be used as positive controls or PCR quantification standards for E. coli O157:H7. This product is compatible with Norgen Biotek’s E. coli O157:H7 Taqman PCR Detection Kit (Cat. TM41350) and E. coli O157:H7 Taqman PCR Detection Probe/Primer and Control Set (Cat. TM41310).

Details

Supporting Data

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Storage Conditions
Upon receipt, store Norgen’s E. coli O157:H7 Quantified Bacterial DNA Standards at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.

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.

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

This product uses an improved salt precipitation purification method to provide a safe and economical solution for High Weight genomic DNA extraction from blood samples, tissue samples, cultured cells, oral swabs, bacteria, and other samples. The extraction does not require the use of toxic phenol chloroform or any expensive reagents, making it the most economical reagent kit for nucleic acid extraction at present. This kit has no limit onthe amount of sample used and can flexibly adjust various amounts of samples. The obtained DNA can be directly used for experiments such as PCR, enzyme digestion, Southern hybridization, and the Third-generation sequencing.

Details

Principles

SolPure DNA Kits is an improved salt precipitation purification method. (Blood samples are lysed in red blood cell lysis buffer to remove red blood cells, and white blood cells are collected by centrifugation.) After lysis, DNA is released into the lysis buffer. RNA is removed by RNASE A. High salt solution is added to precipitate proteins and impurities. Centrifuge to remove precipitate and obtain supernatant containing only DNA. Add isopropanol to precipitate and recover DNA. Wash with 70% ethanol to remove salt, and finally add Buffer TE to dissolve DNA.

Kit Contents

Storage and Stability

RNase A and Proteinase K should be stored at 2–8°C upon arrival. However, short-term storage (up to 4 weeks) at room temperature (15–25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least18 months under these conditions. The entire kit can be stored at 2–8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

This product uses an improved salt precipitation purification method to provide a safe and economical solution for High Weight genomic DNA extraction from blood samples, tissue samples, cultured cells, oral swabs, bacteria, and other samples. The extraction does not require the use of toxic phenol chloroform or any expensive reagents, making it the most economical reagent kit for nucleic acid extraction at present. This kit has no limit onthe amount of sample used and can flexibly adjust various amounts of samples. The obtained DNA can be directly used for experiments such as PCR, enzyme digestion, Southern hybridization, and the Third-generation sequencing.