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Practical 6.2 DNA amplification using PCR

Purpose Safety

• To use PCR to amplify DNA. If you undertake PCR make sure you are aware
of the hazards and follow the instructions of
your teacher very carefully.

Practical PCR
Scientists in forensics laboratories carry out the polymerase chain reaction (PCR) using
a machine called an automated thermal cycler. This is a programmable heating unit in
which the DNA to be amplified is incubated in a buffer solution with thermo-stable DNA
polymerase, primers and deoxyribonucleotides. The unit maintains the cyclical sequence of
temperatures for the PCR process.

Your school or college may be lucky enough to possess a thermal cycler but it is possible to
carry out PCR without them, using three separate thermostatically controlled water baths.
You simply have to move the DNA sample from bath to bath – and complete 30 cycles! You
need a stopwatch, good teamwork and some sort of protection from the steam coming off
the hottest bath.

Having amplified the short tandem repeat sequences within your DNA sample, you will
then separate out the fragments using gel electrophoresis (see Practical 6.1). Comparing
the position of the bands on the gels to a standard or reference you will be able to draw
conclusions about the DNA sample you started with.

You will follow a practical protocol supplied by the company that produces the equipment
and reagents your school or college has purchased.

Edexcel practical materials created by Salters-Nuffield Advanced Biology, ©University of   York Science Education Group.