Kidd Lab 

Arg47His site (located in exon 3)

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

Primer 1. A2FXNFOR:    5'-ATT CTA AAT TGT TTA ATT CAA GAA G-3'       Final concentration 100 ng/25µl
Primer 2. A2FXNREV:    5'-ACT AAC ACA GAA TTA CTG GAC-3'               Final concentration 100 ng/25µl

dNTPs: Final Concentration 200 µM/reaction

Buffer: Final Concentration 2.0 mM MgCl₂, 50 mM KCl, 10 mM Tris-HCl pH 8.4

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 (30"), 56^oC (30'), 72^oC (60")  × 35 cycles
                              72^oC (10')

Product size:
                                 MslI
                   685 bp---------------> 443 bp + 242 bp in homozygote for site present
                                                (site absent is Arg; site present is His)
 
 

Kidd Lab Histograms for Allele Frequencies