The document provides information about polymerase chain reaction (PCR), including its history, definition, applications, and process. It summarizes that PCR was invented in 1984 by Kary Mullis to selectively reproduce portions of DNA through repeated heating and cooling cycles using DNA polymerase. PCR is now an essential tool in biology for applications like detecting genes and diagnosing infections. The process involves extracting DNA, performing PCR cycles of denaturation, primer annealing, and extension, and then analyzing the amplified DNA products through electrophoresis.

1. PCR
Polymerase Chain Reaction
By: Mohammad Adel Rezaei

2. Preface
• Today, molecular biology and genetic
engineering techniques have been widely
used in biological sciences because of the
speed and accuracy.
• These methods can also be used in
diagnostic laboratories.

3. Definition
• a program of periodically repeated heating
and cooling of DNA
• with a heat resistant DNA polymerase
• in order to selective reproduce of a small
portion of the genome
• DNA replication process with this method
is geometrically and therefore large
amounts of DNA can be obtained

4. History
• PCR was invented in 1984 by Kary Mullis
& he received the Nobel Prize in chemistry
in 1993, for his invention.
• is now an essential tool for many
biologists and the standard protocols are
simple and user friendly

5. Applications
• providing multiple copies of a gene
• survey the presence or absence of a
particular gene in a piece of DNA

6. Examples of applications
the prenatal diagnosis of genetic disorders
sex determination
evaluation of viral and bacterial infections

7. Things that should be done
before and after
1. Extracting genetic material
2. PCR process
3. PCR products electrophoresis

8. Extracting genetic material
1. Break the cell
2. Precipitation of DNA and RNA
3. Separation of DNA and RNA
4. Precipitated proteins
5. DNA and RNA isolation from one another
6. Precipitated with ether
7. Spectrophotometry to review and verify
the accuracy of the previous steps

9. The main ingredients for PCR
• Buffer solution
 providing a suitable chemical environment for optimum activity and stability
of the DNA polymerase.
• Four kinds of nucleotides(dNTPs)
• Two primers
• Tag DNA polymerase
• MgCl2
• DNA sample
• Water

10. The main ingredients for PCR
• Buffer solution
• Four kinds of nucleotides(dNTPs)
 the building blocks which the DNA polymerases synthesize a new DNA
strand with them.
• Two primers
• Tag DNA polymerase
• MgCl2
• DNA sample
• Water

11. The main ingredients for PCR
• Buffer solution
• Four kinds of nucleotides(dNTPs)
• Two primers
 perform two operations: First, the location of the gene should be amplified,
and the second to determine the size of self-replicating.
• Tag DNA polymerase
• MgCl2
• DNA sample
• Water

12. The main ingredients for PCR
• Buffer solution
• Four kinds of nucleotides(dNTPs)
• Two primers
• Tag DNA polymerase
 Do DNA replication
• MgCl2
• DNA sample
• Water

13. The main ingredients for PCR
• Buffer solution
• Four kinds of nucleotides(dNTPs)
• Two primers
• Tag DNA polymerase
• MgCl2
 Cofactor for Tag enzyme
• DNA sample
• Water

14. The main ingredients for PCR
• Buffer solution
• Four kinds of nucleotides(dNTPs)
• Two primers
• Tag DNA polymerase
• MgCl2
• DNA sample
 DNA template that contains the DNA region (target) to be amplified.
• Water

15. The main ingredients for PCR
• Buffer solution
• Four kinds of nucleotides(dNTPs)
• Two primers
• Tag DNA polymerase
• MgCl2
• DNA sample
• Water
 As diluent

16. Concentration and volume of
materials for pcr
master mix

17. Concentration and volume of
materials for pcr
Application of PCR in diagnosis of Mycobacterium tuberculosis complex

18. PCR consists of three main steps
• Denaturation
 Double Stranded DNA is denatured by heat into single strands.(94 ° C for 1 minute)
• Primer Annealing
 Primers anneal to the end of the strand.(54 ° C for 45 seconds)
• Primer Extension
 The DNA polymerase recognizes the primer and makes a complementary copy of the
template which is now single stranded.(72 ° C for 2 minutes)

19. PCR three main steps

20. PCR Thermocycler

21. PCR products electrophoresis