Whole-Blood Polymerase Chain Reaction and Restriction Fragment Length Polymorphism: A Simplified Method by Microwave Irradiation

Objectives: The aim of the present study was to develop a simple, quick and cheap method to process whole-blood samples for the molecular techniques polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) without the use of expensive reagents or sophisticated machines. Materials and Methods: Venous whole-blood samples were collected from 40 individuals. The samples were frozen at –80 ° C, and then rapidly thawed at 37 ° C. Each sample was incubated with distilled water, then boiled in a microwave and centrifuged. The supernatant was taken directlyfor PCR and RFLP. For comparison, PCR and RFLP were performed on DNA purified from the same samples using the phenol-chloroform method and two commercial DNA extraction kits. Results: PCR/RFLP results using the presented method were qualitatively similar to those obtained by DNA extracted using the other three methods. Conclusion: The presented method proved to be a simpler and cheaper way of processing whole-blood samples for PCR and RFLP analyses.