Kidd Lab 

PvuII (B) site (located in intron 3)

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

Primer 1.     PAHPVUb-1:    5'-GCC TGA AAG CAA TAC AGA GG-3'        100 ng/reaction
Primer 2.     PAHPVUb-2:    5'-TGA GGT AGG GAT GAA AGA CC-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
                   489 bp ---------------> 259 bp + 230 bp (in homozygote site present)
 
 

Kidd Lab Histograms for Allele Frequencies