Polymerase chain reaction | PCR | class 12

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This lecture is about
The polymerase Chain Reaction
Kary B. Mullis developed the polymerase chain reaction (PCR) in 1983. Earlier methods
of obtaining multiple copies of a specific sequence of DNA were time consuming and
expensive. In contrast, PCR can create millions of copies of a single gene or any specific
piece of DNA quickly in a test tube. PCR is very specific - the targeted DNA sequence
can be less than one part in a million of the total DNA sample. .This means that a single
gene or smaller piece of DNA, among all the human genes can be amplified (copied)
using PCR.
PCR takes its name from DNA polymerase, the enzyme that carries out DNA replication
in a cell. It is considered a chain reaction because DNA polymerase will carry out
replication over and over again, until there are millions of copies of the desired DNA.
PCR does not replace gene cloning, which is still used whenever a large quantity of
gene or protein product is needed.
Before carrying out PCR, primers - sequences of about 20 bases that are complementary
to the bases on either side of the “target DNA” - must be available. The primers are needed
because DNA polymerase does not start the replication process; it only continues or
extends the process. After the primers bind by complementary base pairing to the
DNA strand, DNA polymerase copies the target DNA (Fig 23.5) .
DNA polymerase used is temperature - insensitive (thermostable) enzyme extracted
from the bacterium Thermus aquaticus, which lives in hot springs. Commonly, this
enzyme is also known as Taq polymerase. It can withstand high temperature, which is
used to separate double stranded DNA, therefore, replication need not be interrupted
by the need to add more enzyme. PCR is done these days in an automatic PCR machine
or thermocycler, which is a routine piece of equipment in any laboratory.

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