Team:Newcastle/25 June 2010

Polymerase Chain Reaction protocol

Polymerase chain reaction (PCR) is a technique used to amplify a single or few a copies of a piece of DNA generating thousands to millions of copies of a particular DNA sequence.

1. To avoid contamination, wear gloves
2. To achieve optimum results, always do everything on ice

A basic PCR set up requires several components and reagents.These components include:

1. DNA template that contains the DNA region (target) to be amplified.
2. Two primers that are complementary to the 3' (three prime) ends of each of the sense and anti-sense strand of the DNA target.
3. Taq polymerase or another DNA polymerase with a temperature optimum at around 70 °C.
4. Deoxynucleoside triphosphates, the building blocks from which the DNA polymerases synthesizes a new DNA strand.
5. Buffer solution, providing a suitable chemical environment for optimum activity and stability of the DNA polymerase.