Kidd Lab 

BclI site (located in intron 1, approximately 1 kb upstream from exon 2) (The TaqI (B) and the BclI 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-3'        100 ng/reaction
Primer 2.     RS3BACK1:      5'-GAT ACC CAC TTC AGG AAG TC-3'          100 ng/reaction

dNTPs: Final Concentration 200 µM each

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

Taq DNA Polymerase: 0.5 Units/reaction
                                 (Perkin-Elmer Amplitaq DNA polymerase 5 U/µl, Cat. No. N808-0145)

PCR Cycling Profile: 94^oC (5')
                              94^oC (30"), 60^oC (30"), 72^oC (1')  × 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 TaqI and BclI.

Product size:
                                Bcl I
                   459 bp--------------> 267 bp + 192 bp (in homozygote site present)

Reference:
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.
 
 

Kidd Lab Histograms for Allele Frequencies