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PCR and DNA diagnostics
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PCR AND DNADIAGNOSTICS .
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GROUP :- 3 :1:- HARSH SINGH NIRANJAN2:- ABHISHEK SINGH
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BIOCHEMICAL METHOD:Biochemical analysis techniques refer to a set of methods,assays, and procedures that enable scientists to analyze the
substances found in living organisms and the chemical
reactions underlying life processes.
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POLYMERASE CHAINREACTION:Polymerase chain reaction (PCR) is a method widely used to
rapidly make millions to billions of copies of a specific DNA
sample, allowing scientists to take a very small sample of
DNA and amplify it to a large enough amount to study in
detail.
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TECHNICAL OVERVIEW :Technical overview DNA consists of fourelements: A, C, G and T
DNA molecule :* Double stranded DNA strands.
* Bound together by chemical forces.
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HISTORY OF PCR :*Invented and patented in 1983.*Revolutionary technique.
*Enzymatic DNA amplification.
*Need two short sequences on the DNA.
* Repetition of 30-35 cycles of three steps.
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PCR CYCLE :-8.
The Thepolymerase
chain reaction:polymerase chain reaction
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WHAT U NEED FOR PCR:Two short DNA fragment that stick specifically to each of theDNA strands at some distance of each other Primers
• Can be specific for:°A certain bacterium.
°Bacterial species .
°Genes (e.g., toxin gene).
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WHAT U NEED FOR PCR:Apparatus to perform about temperature procedure• 95 ° 35 C (denaturation of DNA) .
• 5060 ° cycles of a three C (annealing of primers).
• 72 ° C (extension of the primers).
°Sample (+/target DNA).
° Primers for the specific detection.
° Nucleotides .
°Enzyme.
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PERFORMING PCR :1.Put your tube in the apparatus.2:- Let the program run ( 35 cycles).
3:- If primers fit, there is amplification of target DNA.
4:- If primers do not fit, no amplification product => the DNA
(microorganism) was not in the sample
5:-Detect if there is PCR product.
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WHAT IS A DNAFINGERPRINT?
DNA fingerprinting is a
method used to identify an
individual from a sample of
DNA by looking at unique
patterns in their DNA.
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DNA FINGERPRINTING :-14.
QUESTIONS :QUS 1:- What is DNA fingerprinting & Forwhat purpose it is used?
QUS 2:- Describe the steps of PCR ?