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Polymerase Chain Reaction PCR

Polymerase Chain Reaction PCR. What is PCR?. It is a fast and inexpensive technique used to AMPLIFY small fragment of DNA to Produce million of copies. What is PCR used for? Diagnosing acquired or inherited diseases. Diagnosing viral or bacterial diseases. PCR Components. Two Primers

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Polymerase Chain Reaction PCR

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  1. Polymerase Chain ReactionPCR

  2. What is PCR? • It is a fast and inexpensive technique used to AMPLIFYsmall fragment of DNA to Produce million of copies. What is PCR used for? • Diagnosing acquired or inherited diseases. • Diagnosing viral or bacterial diseases.

  3. PCR Components • Two Primers • DNA polymerase • Deoxy-nucleotide triphosphate (dNTP) • Buffer solution • MgCl2 • Extracted DNA sample

  4. PCR Steps: • Denaturation Step: Heating the reaction to 90OC-95OC. • Annealing Step: Rx temperature lowered to 50OC-56OC. • Extention/Elongation: 70OC-72OC. • 30-40 cycles/ Automated using Thermal Cycler. taq

  5. PCR For MTHFR Gene • Objective: To make millions of copies of a specific DNA segment consisting of MTHFR gene. • Materials Given: 3 samples: • Normal • Homozygote • Heterozygote

  6. Materials Given: Master MIX +

  7. PCR Cycles Conditions: 94oC 30 sec 72oC 30 sec 55oC 30 sec 4oC ∞ 33 cycles

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