Polymerase chain reaction, PCR, chemicals used in PCR, Ingredients used in PCR, Deionized water, PCR Buffer, MgCl2, dNTPS, types of dNTPS, Taq DNA polymerase, Primers, Amplicon, Molecular method.

1. CHEMICALS USED IN PCR
BY HASNAT TARIQ

2. CONTENTS
 DEIONIZED WATER
 PCR BUFFER
 MgCL2
 dNTPs
 PRIMERS
 TAQ POLYMERASE
 TARGET DNA SEQUENCE
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3. 1. DEIONIZED WATER
 Also called double distilled water.
 11.4 μl is used per sample in PCR
 Provides physiochemical environment.
 Also dilute chemical reagents.
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4. 2. PCR BUFFER
 10 X buffer is used.
 2μl is used per sample.
 Maintains pH.
 Provides physiochemical
environment.
 Optimize conditions like salt
concentration and pH to enable
Taq polymerase to work
efficiently.
 2 types :
1. KCL
Facilitate template primer
binding.
2. (NH4)2SO4
Increase primer specificity.
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5. 3. MgCL2
 2μl is used per sample.
 Functions as a cofactor of Taq DNA polymerase by providing
magnesium ions.
 Increase polymerase activity of Taq polymerase.
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6. 4. dNTPs
 dNTP stands for
deoxyribonucleotide
triphosphate.
 0.2μl is used per sample.
 There are four different dNTPs.
 The dNTPs serve as the building
blocks for the new strands being
processed during PCR.
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7. 5. PRIMERS
 PCR primers are short fragments of single
stranded DNA (15-30 nucleotides in length)
that are complementary to DNA sequences
that flank the target region of interest.
 The purpose of PCR primers is to provide
a “free” 3'-OH group to which the DNA
polymerase can add dNTPs.
 They are initiators of polymerization.
 Two types : Forward and Reverse Primers
 1μl Forward Primer and 1μl Reverse
primer is used in PCR per sample.
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8. 6. Taq Polymerase
 Obtained from a bacterium
Thermus aquaticus.
 High temperature stability.
 Performs the function of
elongation of DNA strands.
 High rate of dNTP incorporation.
 No proof reading ability.
 0.4μl is used per sample.
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9. 7. TARGET DNA SEQUENCE
 The Target DNA sequence which need to be amplified.
 It is also called amplicon.
 2μl is used per sample.
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10. THANK YOU
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