Description
DRD2 TaqI "B" Polymorphismq

Procedure
Taq I "B" & Bcl I Located in intron 1, approximately 1 kb upstream from exon 2
(The Taq I "B" and the Bcl I sites are 12 bp apart)

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

Primer 1. RS3DWST3 '5- GAT GTG TAG GAA TTA GCC AGG 100 ng/reaction
Primer 2. RS3BACK1 '5- GAT ACC CAC TTC AGG AAG TC 100 ng/reaction

dNTPs Final concentration 200 µM each

Buffer: Final Concentration 1.5 mM MgCl2, 50 mM KCI, 10 mM Tris HCl pH 8.4

Taq DNA Polymerase: 0.5 u/reaction volume of 25 µ1
(Perkin-Elmer Amplitaq DNA polymerase 5u/µl, Cat No. N808-0145)

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

The applicons from these primers encompass two polymorphic sites 12 base pairs apart; if desired divide the product into two and restriction digest with both Taq I and Bcl I

Product size: Taq I
459 bp ——–> 267 bp + 192 bp (in homozogote site present)

Bcl I
459 bp ---------> essentially the same size as with TaqI

Castiglione, C.M., A.S. Deinard, W.C. Speed, G. Sirugo, H.C. Rosenbaum, Y. Zhang, D.K. Grandy, E.L. Grigorenko, B. Bonne-Tamir, A.J. Pakstis, J.R. Kidd, and K.K. Kidd, 1995. Evolution of haplotypes at the DRD2 locus. American Journal of Human Genetics 57:1445-1456.

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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