Optimization of RAPD-PCR conditions in cattle

Random Amplified Polymorphic DNA polymerase chain reaction (RAPD-PCR) is a fast and easy way of identifying DNA polymorphisms generated from several regions of the genome. This could expedite the process of identifying informative polymorphic markers that may be linked to important genes controlling economic traits. In cattle, failure to obtain consistent amplification patterns in RAPD-PCR has been a cause for concern. This has been attributed to the fact that decamer primers that are used in RAPD-PCR reactions are likely to amplify regions of DNA where the primer-template base pairing has some degree of mismatch and that these mismatches fail to repeat from reaction to reaction. This paper describes the use of tricine buffer along with changes in reaction components and thermal cycling conditions that has yielded consistent and reproducible RAPD-PCR amplifications using single primers and double primer combinations on bovine DNA.