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


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.


CONCLUSIONS
The presented method proved to be a simpler and cheaper way of processing whole-blood samples for PCR and RFLP analyses.
Jadaon
Mehrez M
MM
Department of Medical Laboratory Sciences, Faculty of Allied Health Sciences, Kuwait University, Sulaibekhat 90805, Kuwait. mehrez@hsc.edu.kw
Dashti
Ali A
AA
Lewis
Hend L
HL
Habeeb
Fatema M
FM
D054458:Amplified Fragment Length Polymorphism Analysis; D004247:DNA / Q000097:blood / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D006801:Humans; D007962:Leukocytes / Q000737:chemistry; D008872:Microwaves; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length
D004247:DNA