DNA Extraction and Amplification of 10-Day, Room Temperature Blood Samples

WERASAK SASANAKUL, B.Sc.*, SARANYA RURGKHUM, B.Sc. *, PHONGJAN HATHIRAT, M.D.** AMPAIWAN CHUANSUMRIT, M.D.**, UMAPORN UDOMSUBPAYAKUL, M.Sc.*,

Affiliation : * Research Center, ** Department of Pediatrics, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok I 0400, Thailand.

AbstractDNA was serially studied in 20 samples of buffy coat stored at room temperature. Each sample was divided into 5 equal volumes, namely D0, D3, D5, D7 and D10. DNA extraction was performed on days 0, 3, 5, 7 and 10 after blood collection. The mean ratio of OD260/0D280 of the DNA obtained from Do to D10 ranged from 1.77 to 1.79, and the mean amounts of the DNA obtained from Do to D10 ranged from 602 to 740 ng/ul. There were no significant differences in the mean ratio and amounts of DNA obtained among these samples (p >0.05). Subsequently, amplification was successfully performed from this template DNA to yield products of 1.4 kb and 142 bp at the sites associated with B globin and factor VIII genes, respectively. These findings suggest the possibility of sending blood samples for DNA analysis by mail, or no ice is required during transportation.

Keywords : DNA Extraction, Buffy Coat

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