Abi 7900 fast ht sequence detection systems

Genomic DNA samples from 186 unrelated European‐Americans of the San Francisco Bay Area (part of the Studies of Pharmacogenetics in Ethnically Diverse Populations cohort) were obtained and genotyped for the single nucleotide polymorphism rs4149056 (SLCO1B1 c.521T>C). A TaqMan single nucleotide polymorphism genotyping assay (assay ID C__30633906_10) was used for genotyping on the ABI 7900 Fast HT Sequence Detection Systems (Applied Biosystems, Foster City, CA). Genomic DNA (10 ng) was amplified using a TaqMan Genotyping Master Mix (Applied Biosystems). The polymerase chain reaction conditions were as follows: 1 cycle of 95°C for 10 minutes followed by 40 cycles of two‐step polymerase chain reaction with denaturation at 95°C for 15 seconds and annealing and extension at 60°C for 1.5 minutes. Data were analyzed by allele discrimination.

BCRP p.Q141K (rs2231142) was genotyped by TaqMan assay (Applied Biosystems, assay ID C__15854163_70) in DNA extracted from a cheek swab. The reaction mixture consisted of 5–10 ng DNA, 12.5 μl of TaqMan Genotyping Master Mix (Applied Biosystems), 1.25 μl of TaqMan genotyping assay mix, and 11.25 μl of distilled water. The cycling conditions were as follows: 95°C for 10 min and 40 cycles of 95°C for 15 s then 60°C for 1 min. The reaction was run on a BioRad MyCycler (Bio‐Rad) and allele discrimination determined by ABI 7900 Fast HT Sequence Detection Systems (Applied Biosystems).