Genotyping Variants in ANKK1 and DRD2

DNA was isolated from blood using the Gentra Puregene blood kit (Qiagen, Germantown, MD) according to the manufacturer’s recommendations. Genotypes were determined by a researcher who was unaware of the clinical status of the participants using 5′-fluorogenic exonuclease assays (TaqMan®, Applied Biosystems, Foster City, CA). The ANKK1 rs1800497 variant was genotyped using the TaqMan® primer-probe sets (Applied Biosystems) assay ID C_7486676_10, and the DRD2 rs2283265 variant with assay ID C__16070796_10. PCR amplifications were performed in duplicate using Platinum® quantitative PCR SuperMix-UDG (Invitrogen, Carlsbad, CA) on a ViiA7 and ViiA 7 Software v1.1 was used for data analysis (Applied Biosystems). SRY was genotyped to determine sex (Kosten et al., 2013 (link)). Ten ancestry-informative markers were genotyped to determine population structure, and the data from our cohort were compared against CEPH-HGDP samples (1,035 subjects of 51 populations) as previously described (Kosten et al., 2013 (link)).

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Spellicy C.J., Harding M.J., Hamon S.C., Mahoney JJ I.I.I., Reyes J.A., Kosten T.R., Newton T.F., De La Garza R I.I, & Nielsen D.A. (2014). A variant in ANKK1 modulates acute subjective effects of cocaine: a preliminary study. Genes, brain, and behavior, 13(6), 559-564.

Publication 2014

Gentra Puregene blood kit TaqMan® primer-probe sets Platinum® quantitative PCR SuperMix-UDG ViiA 7 Software v1.1

5 exonuclease Assay Blood Drd2 Platinum Populations Primer

Corresponding Organization :

Other organizations : Michael E. DeBakey VA Medical Center, Baylor College of Medicine, Rockefeller University, Rice University

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Protocol cited in 2 other protocols

Variable analysis

independent variables

ANKK1 rs1800497 variant
DRD2 rs2283265 variant

dependent variables

Genotypes

control variables

Clinical status of the participants
Population structure

controls

Positive control: SRY genotyping to determine sex
Negative control: Comparison of population structure data against CEPH-HGDP samples (1,035 subjects of 51 populations)

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