Kidd Lab 

BglII site (located at 5' end of exon 1)

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

Primer 1.     B2PAH1:    5'-GCA GGA AAC TCT CTG ACT TTG G-3'        100 ng/reaction
Primer 2.     B2PAH2:    5'-TGG CAG TTC TGG AGG CCA GA-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"), 60^oC (30"), 72^oC (30")  × 30 cycles

Product size:
                                BglII
                   290 bp ---------------> 209 bp + 81 bp (in homozygote site present)
 
 

Kidd Lab Histograms for Allele Frequencies