Kidd Lab 

HindIII (A) site (located 14 kb upstream (telomeric) of HindIII (B) site)

Template: Genomic DNA 100-200 ng/reaction volume of 20 µl

Primer 1.     PKOH3AF:     5'-CCA TAA CAA TTA CAG AGA ATT GAG-3'            10 pM/reaction
Primer 2.     PKOH3AR:     5'-CCC TCA GGG AGG CAG CCC-3'                         10 pM/reaction

dNTPs: Final Concentration 200 µM/reaction

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"), 60^oC (30"), 72^oC (30")  × 30 cycles

Product size:
                                 HindIII
                    234 bp---------------> 194 bp and invisible 40 bp for homozygote site present

Reference: Mclntosh et al., 1989. Amer. J. Med. Genet. 32:274-276.
 
 

Kidd Lab Histograms for Allele Frequencies