POLYMERASE

CHAIN REACTION
Presented by : Cyril C. Castillo
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WHAT IS
POLYMERASE CHAIN
REACTION?
Polymerase chain reaction (PCR) is a
common laboratory technique used to
make many copies of a particular region
of DNA.
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PCR HAS FIVE CORE

‘INGREDIENTS’:
•the DNA template to be copied.
•primers – short stretches of DNA or RNA, 20 to 30
bases in length, that bind either side of the DNA
section of interest and mark the point that PCR starts.
•DNA nucleotide bases (also known as dNTPs). DNA
bases (A, T, C and G) are the building blocks of DNA
and are needed to construct the new strand of DNA.
•an enzyme, called Taq polymerase, which adds the
bases to the copied sequence.
•a buffer to ensure the right conditions for the reaction.
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TAQ POLYMERASE
PCR requires a DNA polymerase
enzyme that makes new strands of
DNA, using existing strands as
templates. The DNA polymerase
typically used in PCR is
called Taq polymerase, after the heat-
tolerant bacterium from which it was
isolated (Thermus aquaticus).
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PCR PRIMERS
Taq polymerase can only make DNA if it's
given a primer, a short sequence of
nucleotides that provides a starting point for
DNA synthesis. PCR primers are short
pieces of single-stranded DNA, usually
around nucleotides in length. Two primers
are used in each PCR reaction, and they are
designed so that they flank the target region
(region that should be copied).
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STEPS IN PCR
The basic steps are: 7

1.Denaturation : Heat the reaction strongly to

separate, or denature, the DNA strands. This
provides single-stranded template for the next
step.
2.Annealing : Cool the reaction so the
primers can bind to their complementary
sequences on the single-stranded template
DNA.
3.Extension : Raise the reaction temperatures
so Taq polymerase extends the primers,
synthesizing new strands of DNA.
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GEL ELECTROPHORESIS
The results of a PCR reaction are
usually visualized (made visible)
using gel electrophoresis, it is a
technique in which fragments of
DNA are pulled through a gel
matrix by an electric current, and it
separates DNA fragments
according to size.
FIELDS WHERE PCR
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IS USED
• Medical diagnostics
• Forensic science
• Research
• Food safety
• Environmental science
• Pharmaceuticals
• Veterinary medicine
• Plant biology
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IMPORTANCE OF
PCR

PCR has become an indispensable

technology with diverse applications
that has impacted our health, safety,
environment and the overall quality
of life.
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QUIZ TIME
 Primers Denaturation 13
Annealing Taq Polymerase
Extension Polymerase chain reaction

1. Common laboratory technique used to make

many copies of a particular region of DNA.
2. A short sequence of nucleotides that provides a
starting point for DNA synthesis
3. The DNA polymerase typically used in PCR
4-6. Steps in PCR
7. Named after the heat-tolerant bacterium from
which it was isolated (Thermus aquaticus).
8-10. 3 fields where pcr is used
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