Kidd Lab 

Arg369Cys site (located in exon 9)

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

Primer 1. HE39:    5'-TGG ACT TCA CAA CAA GCA TGT-3'               Final concentration 100 ng/25µl
Primer 2. HE40:    5'-TTG ATA ACA TCT CTG AAG AGC TGA-3'        Final concentration 100 ng/25µl

dNTPs: Final Concentration 200 µM/reaction

Buffer: Final Concentration 1.5 mM MgCl₂, 50 mM KCl, 10 mM Tris-HCl pH 8.4
DMSO: Final Concentration 5%

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

PCR Cycling Profile: 95^oC (5')
                              95^oC (15"), 58^oC (15"), 72^oC (60")  × 40 cycles
                              72^oC (10')

Product size:
                                 AlwNI
                   201 bp ---------------> 130 bp + 71 bp in homozygote site present
                                                (site absent is Arg; site present is Cys)

Reference:  Xu et al. 1998. Genotyping of human alcohol dehydrogenases at the ADH2 and ADH3 loci following DNA sequence amplification. Genomics 2:209-214.
 
 

Kidd Lab Histograms for Allele Frequencies