Kidd Lab 

Sau3A site (RET4) (only rare heterozygotes in Biaka and Europeans)

Template: Genomic DNA 50-100 ng/reaction volume of 25 µl

Primer 1.     S94Sau3F2:        5'-TTG GGA CAG TGA TGA GGA ACT-3'          100 ng/reaction
Primer 2.     RETRESQ4R:     5'-CCC TCC ATA CAG CAC GGT TCT-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: 95^oC (5')
                              95^oC (30"), 58^oC (30"), 72^oC (1')  × 30 cycles

Product size:
                                 Sau 3A
                   625 bp ------------------>  411 bp +  214 bp(constant band) in site abesent
                                 Sau 3A
                   625 bp ------------------->  335 bp +  76 bp + 214 bp(constant band) in site present
 
 

Kidd Lab Histograms for Allele Frequencies