PCRmax Bacillus anthracis Instruction Manual

Hypothetical protein
Bacillus anthracis
PCRmax Ltd qPCR test
TM
For general laboratory and research use only
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Bacillus anthracis is a Gram-positive, rod-shaped, non-motile, spore bearing bacterium with a circular DNA genome of around 5.2Mbp with plasmids. B. anthracis is the only obligate pathogen within the Bacillus genus and causes Anthrax, an acute lethal infection of both ruminant animals and humans.
B. anthracis spores are soil borne and can survive for decades even under extreme and unfavourable conditions. Spores enter a host by inhalation, ingestion or via skin lesions. Subsequently they penetrate the blood stream, germinate and produce fatal toxins. pXO2 encodes the polyglutamyl capsule that [protects the organism from host phagocytosis. pXO1 plasmid encodes three proteins that in complex, form a toxin although two of these components are toxic in their own right: EF (oedema factor) increases cyclic AMP levels causing oedema while LF (Lethal factor) destroys white blood cells. PA (protective antigen) facilitates cell entry of the other two proteins by pore formation.
Cutaneous anthrax ,caused via spore contact with skin lesions, is the most common form in humans accounting for 90% of all reported cases and rarely fatal if treated. This form initially presents as a boil-like skin lesion between 2-3 days after exposure and eventually forms a painless ulcer with a black center (eschar). Individuals with the ingested, gastro­intestinal form undergo a similar disease progression but mortality rates are much higher. Higher still is the mortality rate of inhaled, pulmonary anthrax. Macrophages in the alveolar provide entry to the blood stream which can result in septic shock and respiratory difficulties progressing to organ failure.
Anthrax is generally treated with antibiotics with penicillin being the drug of choice with current vaccination offering only temporary protection.
Introduction to Bacillus anthracis
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MIN
MAX
The PCR Max qPCR Kit for Bacillus anthracis (B.anthracis) genomes is designed for the in vitro quantification of B.anthracis genomes. The kit is designed to have the broadest detection profile possible whilst remaining specific to the B.anthracis genome.
The primers and probe sequences in this kit have 100% homology with a broad range of B.anthracis sequences based on a comprehensive bioinformatics analysis.
If you require further information, or have a specific question about the detection profile of this kit then please send an e.mail to help@pcrmax.com and our bioinformatics team will answer your question.
Specificity
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Kit Contents
• B.anthracis specific primer/probe mix (150 reactions BROWN)
FAM labelled
• B.anthracis positive control template (for Standard curve RED)
• Internal extraction control primer/probe mix (150 reactions BROWN)
VIC labelled as standard
• Internal extraction control DNA (150 reactions BLUE)
• Endogenous control primer/probe mix (150 reactions BROWN)
FAM labelled
• RNAse/DNAse free water (WHITE)
for resuspension of primer/probe mixes and internal extraction control DNA
• Template preparation buffer (YELLOW)
for resuspension of positive control template and standard curve preparation
Reagents and equipment to be supplied by the user
Real-Time PCR Instrument
DNA extraction kit
This kit designed to work well with all processes that yield high quality DNA with minimal PCR inhibitors.
Lyophilised 2x qPCR Mastermix
This kit is designed to be compatible with all commercially available Mastermixes that run with standard cycling conditions.
Pipettors and Tips
Vortex and centrifuge
Thin walled 1.5 ml PCR reaction tubes
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