# Polymerase chain reaction # Introduction of PCR # Application of PCR # Schematic drawing of a complete PCR cycle # Thermal cycling :majority of PCR methods rely # Principle of PCR # ONE STEP & TWO STEP RT--PCR # PCR : 03 STEP # PCR : Process diagram

1. Polymerase chain reaction
• Presented by :
Khushbu’s Microbiology

2. INTRODUCTION
• Polymerase chain reaction (PCR) :
• method widely used to rapidly
make millions to billions of copies
(complete copies or partial copies)
of a specific DNA sample,
allowing scientists to take a
very small sample of DNA and
amplify it (or a part of it) to a
large enough amount to study in detail.

3. INTRODUCTION
• Polymerase chain reaction (PCR) :
method widely used to
rapidly make millions to billions of
copies (complete copies or partial
copies) of a specific DNA sample,
allowing scientists to take a
very small sample of DNA &
amplify it (or a part of it) to a
large enough amount to study
in detail.
• Image : A strip of eight PCR tubes, each containing a
100 μL reaction mixture

4. Schematic drawing of a complete PCR cycle

5. • Thermal cycling:
Thermal cycling :majority of PCR methods rely :
• Exposes reactants to repeated cycles of heating
and cooling to permit different temperature-
dependent reactions – specifically,
DNA melting
enzyme-driven DNA replication.
• PCR employs two main reagents
1 primers
2. DNA polymerase.
• IMAGE :A modern
high-throughput thermal cycler

6. Principles
PCR amplifies a specific region of a DNA strand
(the DNA target).
 Most PCR methods amplify DNA fragments of
between 0.1 and 10 kilo base pairs in length,
although some techniques allow for
amplification of fragments up to 40 kbp.
 The amount of amplified product is
determined by the available substrates in the
reaction, which becomes limiting as the
reaction progresses

7. ONE STEP & TWO STEP RT+PCR

8. PCR :STEP