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

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

Factor V Kuwait: a novel mutation in the coagulation factor V gene discovered in Kuwait

It was the aim of this study to report a new point mutation in the clotting factor V gene in the general Arab population. The HR2 haplotype was tested in 288 Arabs living in Kuwait - 188 patients with venous thromboembolic disorders [VTE] and 100 healthy subjects - using polymerase chain reaction and restriction fragment length polymorphism techniques. The presence of the new mutation was verified by DNA sequencing. Two [1.06%] VTE patients had guanine instead of the wild-type adenine at nucleotide number 3935 [A3935G] of the factor V gene. This mutation caused a histidine to arginine change in amino acid number 1254 of the factor V molecule. The new mutation is termed 'factor V Kuwait' [His1254Arg] and was absent in the 100 healthy subjects. It appears that factor V Kuwait could be a risk factor for developing VTE in Arabs. A larger study is needed to confirm this observation