DNA PCR Sequencing

In vitro amplification of a DNA sequence can be achieved by Polymerase Chain Reaction or PCR. The requirements for a PCR are, the DNA template strand to be duplicated, specific DNA primers, nucleotide bases, DNA Taq polymerase and a thermocycler in which the reaction will occur. The reaction consists of the following steps: 1. The DNA sequence to be amplified is identified. 2. DNA primers are designed such that they provide 3' ends for replication of the template using DNA polymerase. 3. The DNA to be amplified is heated to cause strand separation. 4. The system is cooled to permit addition and adhesion of the primers to the respective strands. 5. Hybridization of the DNA occurs in the presence of DNA polymerase and the nucleotide bases. At the end of cycle 2 molecules of DNA are obtained from a single DNA molecule. A detailed explanation of the process and an animated version can be seen in the following videos.