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

Description

The ExcelTaq™ 5X Blood Direct PCR Master Mix Kit is designed for amplifying targeted DNA directly from whole blood, eliminating the need for a lengthy DNA isolation process. ExcelTaq™ Blood Direct PCR Master Mix Kit is a unique blend of 5X concentrated mixture containing all the essential components of a PCR reaction. The ExcelTaq™ Blood Direct PCR Master Mix Kit also contains the PCR-friendly loading and tracking dye solution Orange G. The inclusion of Orange G tracking dye allows for a more convenient post-PCR gel electrophoresis analysis. ExcelTaq™ 5X Blood Direct PCR Master Mix Kit is capable of tolerating the presence of PCR interfering/ inhibiting substances in blood. The ExcelTaq™ 5X Blood Direct PCR Master Mix Kit is ideal for high-throughput screening of blood samples for high reproducibility. ExcelTaq™ 5X Blood Direct DNA PCR Master Mix Kit includes a pair of Positive Control Primers (CCR5) that are compatible with primate blood samples. 

Features

• 5’→3′ DNA polymerase activity
• No detectable 3’→5′ exonuclease (proofreading) activity
• Generates PCR products with 3′-dA overhangs 
• High throughput PCR
• High yield PCR 
• High reproducibility, with reduced pipetting errors
• Compatible with most anticoagulants

Applications 

• Direct amplification of DNA from blood samples
• High throughput screening without DNA purification 
• Suitable for multiplex PCR

Storage

Caution: Avoid Multiple Freeze/Thaw Cycles

4°C for 6 months
-20°C for 24 months

The ExcelTaq™ 5X Blood Direct PCR Master Mix Kit is designed for amplifying targeted DNA directly from whole blood, eliminating the need for a lengthy DNA isolation process. ExcelTaq™ Blood Direct PCR Master Mix Kit is a unique blend of 5X concentrated mixture containing all the essential components of a PCR reaction. The ExcelTaq™ Blood Direct PCR Master Mix Kit also contains the PCR-friendly loading and tracking dye solution Orange G. The inclusion of Orange G tracking dye allows for a more convenient post-PCR gel electrophoresis analysis. ExcelTaq™ 5X Blood Direct PCR Master Mix Kit is capable of tolerating the presence of PCR interfering/ inhibiting substances in blood. The ExcelTaq™ 5X Blood Direct PCR Master Mix Kit is ideal for high-throughput screening of blood samples for high reproducibility. ExcelTaq™ 5X Blood Direct DNA PCR Master Mix Kit includes a pair of Positive Control Primers (CCR5) that are compatible with primate blood samples.

Introduction

This Kit is designed to purify genomic DNA and total RNA simultaneously from a single biological sample. Lysate is first passed through a DNA spin column to selectively isolate DNA and then through an RNA column to selectively isolate RNA. Pure DNA and RNA are purified from the entire sample, in contrast to other procedures where either the biological sample or the purified total nucleic acids is divided into two before being processed separately. The kit is compatible with small amounts of a wide range of animal cells and tissues.

Details

Specifications

Features	Specifications
Main Functions	Co-isolation DNA and RNA from skin, muscle, connective tissue, fibrous tissue sample
Applications	PCR and southern blot, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Cultured cells and tissue samples
Sample amount	Tissue: < 20mg

Principle

The Kits are designed to purify both genomic DNA and total RNA from the same cellor tissue sample. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column and bind DNA. Ethanol is added to the flow-through and the sample is applied to an RNA column. DNA/RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30µl water using the Kit. High-quality DNA is eluted in as little as 50µl water using the Kit.

Advantages

• High quality – high purity total RNA / DNA can be directly used in a variety of downstream applications
• Fast – column method can complete the extraction of several samples in 30 minutes
• Safe – no phenol chloroform extraction required
• Simultaneous extraction- simultaneously isolate DNA and RNA from one sample

Kit Contents

Contents	R511402	R511403
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Columns	50	250
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	100	2 x 250
Buffer RL	30 ml	150 ml
RNA Digestion Buffer	15 ml	80 ml
Buffer DW1	30 ml	150 ml
Buffer RW1	50 ml	200 ml
Buffer RW2*	20 ml	2 x 50  ml
RNase Free Water	10 ml	30 ml
Buffer AE	10 ml	50 ml

Storage and Stability

HiPure DNA/RNA Kit can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. Make sure that all buffers are at room temperature when used. During shipment, crystals or precipitation may form in the Buffer RLC. Dissolve by warming buffer to 37°C.

This Kit is designed to purify genomic DNA and total RNA simultaneously from a single biological sample. Lysate is first passed through a DNA spin column to selectively isolate DNA and then through an RNA column to selectively isolate RNA. Pure DNA and RNA are purified from the entire sample, in contrast to other procedures where either the biological sample or the purified total nucleic acids is divided into two before being processed separately. The kit is compatible with small amounts of a wide range of animal cells and tissues.