Published Online: 1 November 2003
Page Count: 2
Genomic DNA was isolated from buccal swabs by the QIAamp DNA Mini Kit (QIAGEN). The primers used for amplification and sequencing of exon 6 and 7 of ABO gene were suggested by Lee and Chang (1), and Ogasawara et al. (2), respectively. About 1-2 ng of genomic DNA was used for PCR in 25 μL reaction volume. PCR mixture contained 10 mM Tris-HCl pH 8.5, 50 mM KCl, 1.5 mM MgCl2, 200 μM of each dNTP, 0.25 μM of each primer, and 1 U of AmpliTaq Gold DNA Polymerase (Applied Biosystems). Thermal cycling was performed initially at 95°C for 10 min, then 35 cycles consisting of 95°C for 1 min, 60°C for 1 min for exon 6 and 63°C for 1 min for exon 7, 72°C for 1 min, followed by 10 min extension at 72°C in a GeneAmp PCR System 9600 (Perkin Elmer).
Paper ID: JFS2003169