PCR

General Information

PCR is an acronym for polymerase chain reaction. It is a method for amplifying DNA in vitro.

The Basic PCR protocol calls for first heating double stranded DNA at 95 degrees celcius to separate the strands. This is followed by annealing PCR primers (55 celcius) to the single stranded DNA at the 5' ends of the DNA. A specialized enzyme, TAQ polymerase, is then added to the reaction mixture at 72 celcius (TAQ's optimum temperature). TAQ polymerase will use the primers to synthesize new DNA, creating copies of the original strand. The process is repeated numerous times for large amplification results.

Specific Protocols

• Silver: PCR
• E. huxleyi PCR
• Knight:Colony PCR

Notes

1. A discussion of the amplification efficiencies of different DNA polymerases on templates of varying length and GC content using real-time PCR [1].

References

1. Arezi B, Xing W, Sorge JA, and Hogrefe HH. Amplification efficiency of thermostable DNA polymerases. Anal Biochem. 2003 Oct 15;321(2):226-35. DOI:10.1016/s0003-2697(03)00465-2 |