POLYMERASE CHAIN REACTION (PCR) AND APPLICATIONS(1)

NATIONAL INSTITUTE OF HEMATOLOGY AND BLOOD TRANSFUSION POLYMERASE CHAIN REACTION (PCR) AND APPLICATIONS Genetic and Molecular Biology Department Nguyen Thi Quynh Contents Overview of PCR Steps of PCR Applications of PCR Overview Polymerase Chain Reaction (PCR)  The PCR is a popular technique in molecular biology to amplify DNA fragment without using living organisms such as E coli or fungus  The PCR is used in biological and medical studies  The PCR method is invented in 1985 by Kary Mullis and his colleagues (American) Principle Principle Principle Step Step Step Denaturation Annealing Extension Principle Components of a PCR reaction Steps of PCR Step 1: Denaturation At 94 ° C, Double-stranded DNA melts → single-stranded DNA Steps of PCR Step 2: Annealing At a temperature of about 54°C primers bind to single-stranded DNA (template) Step of PCR Step 3: Extension At 72 ° C, enzyme Tag polymerase elongates the complementary sequence, using dNTPs Applications of PCR  Screening of human DNA samples for mutations related to genetic diseases such as: Thalassemia, Hemophilia…  Can detect and quantify the presence of the virus such as: HBV, HCV, CMV…  Detecting gene mutations  Etc Applications of PCR  Tests are performed by PCR technique in Genetic and Molecular biology Department:      Leukemia genes Thalassemia genes Hemophilia genes Prenatal diagnosis in Thalassemia and Hemophilia Quantification of Virus: HBV, HCV, CMV THANKS FOR YOUR ATTENTION!!! ...Contents Overview of PCR Steps of PCR Applications of PCR Overview Polymerase Chain Reaction (PCR)  The PCR is a popular technique in molecular biology to amplify DNA fragment... reaction Steps of PCR Step 1: Denaturation At 94 ° C, Double-stranded DNA melts → single-stranded DNA Steps of PCR Step 2: Annealing At a temperature of about 54°C primers bind to single-stranded... organisms such as E coli or fungus  The PCR is used in biological and medical studies  The PCR method is invented in 1985 by Kary Mullis and his colleagues (American) Principle Principle Principle