PCR method of amplying proteins
The polymerase chain reaction(PCR) is used to amplify specific DNA or RNA sequences through an enzymatic process. It requires upstream and downstream primers, DNA polymerase, four dNTP’s (dATP, dCTP, dGTP, and dTTP) a magnesium ion and a DNA template to be amplified. PCR is performed in three steps. In the first step, the DNA is heated to 95°C where it becomes denatured. In the second step, the DNA is cooled to 55°C for annealing of the primer. The two primers line up complimentary to the 3’ starting point of the sequence. These primers must be long enough to bind to a unique site on the template DNA. Once the DNA has lined up and annealed with the primer, the DNA is reheated in the final step to 72°C. This allows the DNA polymerase to elongate and amplify the target sequence. This cycle is repeated, producing two new fragments of product duplex DNA per template DNA.
