Kidd Lab 

RsaI site(located in intron 3)

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

Primer 1.     HRKin03F4:     5'-TGC CTC AGC CTC CCC ACT A-3'               100 ng/Reaction
Primer 2.     HRKIN03R:      5'-ACC ATG TCC AAA TCC ACC AG-3'            100 ng/Reaction

dNTPs: Final Concentration 250 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 (1')  × 35 cycles
                              72^oC (10')

Product size:
                                     RsaI
                    606 bp----------------> 347 bp + 259 bp (in homozygote site present)
 
 

Kidd Lab Histograms for Allele Frequencies