POLYMERASE CHAIN REACTION (PCR) AND APPLICATIONS(1)

POLYMERASE CHAIN REACTION PCR AND APPLICATIONSGenetic and Molecular Biology Department Nguyen Thi Quynh NATIONAL INSTITUTE OF HEMATOLOGY AND BLOOD TRANSFUSION... Overview of PCRSteps of

POLYMERASE CHAIN REACTION (PCR) AND APPLICATIONS

Genetic and Molecular Biology Department

Nguyen Thi Quynh

NATIONAL INSTITUTE OF HEMATOLOGY AND BLOOD TRANSFUSION

Overview of PCR

Steps of PCR

Applications of PCR Contents

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

Step 1

Denaturation

Step 2

Annealing

Step 3 Extension

Principle

Components of a PCR reaction

Principle

Steps of PCR

At 94 ° C, Double-stranded DNA melts → single-stranded DNA

Step 1: Denaturation

Steps of PCR

At a temperature of about 54°C primers bind to single-stranded DNA (template)

Step 2: Annealing

Step of PCR

At 72 ° C, enzyme Tag polymerase elongates the complementary sequence, using dNTPs.

Step 3: Extension

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

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