Kidd Lab 

TEMPLATE: Genomic DNA 100ng/reaction volume of 25 µl

PRIMER 1.  A2IN3DW3     5'  ATA TTT ATT TTA CCC TAA ACT TAT G 3'
PRIMER 2.  A2IN3UP2      5'  GAG CTA AAA CAT ACT TTG GAT AG 3'

10 µM each primer

dNTPs: Final concentration 200µM

BUFFER: Final concentration 2.2mM MgCl₂, 50mM KCl, 10mM Tris HCl pH 8.4

TAQ DNA Polymerase: 1unit (Perkin-Elmer Amplitaq DNA polymerase 5u/µ l, Cat No. N808-0145)

PCR Cycling Profile: 94^oC (5')
                               94^oC (40)", 59^oC (30"), 72^oC (1.0')  × 30 cycles
                               72^oC (10')
  product size:
                   RsaI
     297 bp --------------> 178 bp and 119 bp (in site present homozygote)

NOTE: Partial intron3 sequence of ADH2 is in GenBank accession #AF040967. The polymorphic nucleotide is 790 bp downstream of exon3 at position 956 of that GenBank entry.

Protocol modified from a RFLP protocol described by Moyra Smith ("Genetics of human alcohol and aldehyde dehydrogenases" Adv. Hum. Genet. 15:249-290, 1986).
 

Kidd Lab Histograms for Allele Frequencies