PCR Cloning

Polymerase Chain Reaction (PCR)
- it is the direct method of making copies of a desired DNA sequence
- it is similar to DNA replication in a way that they both occur in the nucleus
- heat (94°C ~ 96°C) is used to seperate the hydrogen bonds between complementary bases, thus creating two template strands
- temperature is then cooled off to about between (50°C ~ 65°C) to allow the primers to anneal
- DNA primers are used instead of RNA primers and attach to the 3' end of the gene of interest
- two primers (forward and reverse) are extended by using Taq polymerase after heating to 72°C
- Taq polymerase then synthesizes the complementary strands from free nucleotides
- steps are repeated for the two newly created sets of semi-conservative DNA

For clearer resolution of the iamge below click on http://www2.le.ac.uk/departments/emfpu/genetics/explained/images/PCR-process.gif