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Polymerase Chain Reaction

Polymerase chain reaction (PCR) is a technique used to amplify a specific region of DNA through repeated cycles of heating and cooling. It involves using primers that are complementary to the target DNA region, along with a DNA polymerase and free nucleotides. During each cycle, the DNA is denatured, the primers anneal to the DNA, and the polymerase extends the DNA strands, doubling the amount with each round. After 30 rounds, there are over a billion copies of the target DNA region that can then be analyzed.

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345 views

Polymerase Chain Reaction

Polymerase chain reaction (PCR) is a technique used to amplify a specific region of DNA through repeated cycles of heating and cooling. It involves using primers that are complementary to the target DNA region, along with a DNA polymerase and free nucleotides. During each cycle, the DNA is denatured, the primers anneal to the DNA, and the polymerase extends the DNA strands, doubling the amount with each round. After 30 rounds, there are over a billion copies of the target DNA region that can then be analyzed.

Uploaded by

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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Polymerase Chain Reaction

- Template DNA

Reagents:

(containing the gene to be amplified)


-

Primers

Primers are small RNA sequences that are


recognized by the polymerase,
complimentary to the beginning and end of
the gene.
-

Taq polymerase
- free nucleotides

Polymerase Chain Reaction

Step 1: Denature
The tube containing all the
necessary reagents is heated
to ~90C (194F).
At this temperature, the
hydrogen bonds holding the
template DNA strands
together are broken.

Polymerase Chain Reaction


Step 2: Anneal
The tube is cooled to 50C
(122F).
At this temperature, the two
primers bind to their
complimentary regions of the
template DNA.

Step 3: Elongation
The tube is heated to 72C
(161F).
At this temperature, Taq
polymerase binds to the
primers and beginsTaqelongating
the DNA strand, using the free
dNTPs.

At the end of 1 round of


PCR, we have two DNA
strands that contain the
gene of interest.
We now repeat ~30
times, until we have
enough DNA to analyze.

Round 1:
2 DNA Copies

Round 2:
4 DNA Copies

2 copies of
the target
gene!

Round 3:
8 DNA Copies

Polymerase Chain
Reaction
For n rounds of PCR, there are 2n DNA copies made for each
template DNA.
After 30 round of PCR, there are ~1.07 billion copies of the gene.

If primers are carefully chosen to amplify only the gene of


interest, the result will be very pure.

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