Kidd Lab 

TaqI site (located about in the middle of the pKO-82 clone)

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

Primer 1.     PKO82CW5:      5'-CTG CTG GGC ATC GGA GTG-3'                10 pM/reaction
Primer 2.     PKO82RCK2:     5'-CAC AGG AGA AAG GAG GGA GC-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"), 59^oC (30"), 72^oC (30")  × 30 cycles

Product size:
                                 TaqI
                     310 bp--------------> 200 bp + 110 bp (in homozygote site present)

Reference: TaqI RFLP site located in Kidd lab by sequencing.
 
 

Kidd Lab Histograms for Allele Frequencies