Kidd Lab 

PvuII(A) site (located near the 5' end of intron 2)

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

Primer 1.     PAHPVU2-1:    5'-GGC ATG ACT GGA TAC GAT TAG-3'        100 ng/reaction
Primer 2.     PAHPVU2-2:    5'-CTA GAC TCA GAA TGC CTG GG-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"), 58^oC (30"), 72^oC (30")  × 30 cycles

Product size:
                                 PvuII
                   374 bp ---------------> 225 bp + 149 bp (in homozygote site present)
 
 

Kidd Lab Histograms for Allele Frequencies