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 degrees C, and then rapidly thawed at 37 degrees C. Each sample was incubated with distilled water, then boiled in a microwave and centrifuged. The supernatant was taken directly for 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.