Whole-blood polymerase chain reaction and restriction fragment length polymorphism: a simplified method by microwave irradiation

ABSTRACT

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. 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 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. PCR/RFLP results using the presented method were qualitatively similar to those obtained by DNA extracted using the other three methods. The presented method proved to be a simpler and cheaper way of processing whole-blood samples for PCR and RFLP analyses