DNA Amplification by Polymerase Chain Reaction PCR

Random 10bp primers used were A05 (= Set A, primer 05), A05 = 5 AGGGGTCTTG3, and All, CAATCGCCGT, both purchased from Operon Technologies, Alameda, California. Nucleotides (dNTPs) were from both Promega (Madison, WI, USA) and Perkin-Elmer Cetus, Norwalk, CT, USA). Taq polymerase was AmpliTaq (DNA polymerase AS), from Perkin-Elmer Cetus. The DNA size marker was either the 100-bp ladder or the 123-bp ladder, both were purchased from Gibco (Gaithersburg, MD, USA). 

The reliability of the PCR (polymerase chain reaction) is dependent on many external factors, such as false-negative results caused by uncontrolled PCR inhibitor, but also by contamination with foreign DNA. Special attention has to be given to the breakdown of the PCR by contamination through the air the contamination 

Protocol for the amplification of chestnut DNA The reaction mixture is vortexed and covered with a drop of mineral oil in order to reduce evaporation during the amplification process. The tubes are put in the aperture of the thermocycler, which is in advance filled with a drop of mineral oil in order to guarantee close contact between the wall of the cycler and the tube. 

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Describe the procedure for DNA amplification by polymerase chain reaction (PCR). 

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