Description
DRD2 TaqI "D" Polymorphism

Procedure
Taq I "D" Located approximately 3.5 kb downstream from exon 2

Template: Genomic DNA 100-200 ng/reaction volume of 25 µl

Primer 1: TaqDck4 '5 CCT CTG AGG CTT ACT GTC TG 3' 100ng/Reaction
Primer 2: D2Ex2UP7 '5 AAA ACT AGG GAG GGT CAG AG 3' 100 ng/Reaction

dNTPs: Final concentration 200 µM each

Buffer: Final concentration 1.5 mM MgCl2. 50 mM KCl, 10 mM Tris HCI pH 8.4

Taq DNA Polymerase: 0.5u/reaction volume of 25 µI
(Perkin-Elmer Amplitaq DNA polymerase 5u/µI, Cat No. N808-0145)

PCR Cycling Protocol 94oC (5')
94oC ( 30"), 60oC ( 30"), 72oC ( 1.0') × 30 cycles

These primers recognize two polymorphisms just 160 bp apart; if desired divide the product in two and digest with both enzymes

Product size: TaqI
300 bp ——–> 212 bp + 88 bp (homozygote site present)


Kidd, K.K., B. Morar, C.M. Castiglione, H. Zhao, A.J. Pakstis, W.C. Speed, B. Bonne-Tamir, R.-B. Lu, D. Goldman, C. Lee, Y.S. Nam, D.K. Grandy, T. Jenkins, and J.R. Kidd, 1998. A global survey of haplotype frequencies and linkage disequilibrium at the DRD2 locus. Human Genetics 103:211-227.

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