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Understanding PCR Process: A Powerful DNA Amplification Technique

PCR is a rapid, cost-effective method to replicate small DNA fragments, crucial for diagnosing diseases and identifying genetic traits. Learn about its components, steps, and application in gene amplification.

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Understanding PCR Process: A Powerful DNA Amplification Technique

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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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