Kidd Lab 

StyI site (located in intron 5)

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

Primer 1. A7IN5DW2:    5'-TAT TAA ATT ATT GCT TAA TAA CTG G-3'       Final concentration 100 ng/25µl
Primer 2. A7IN5UP1:     5'-TTC CTG TGT CTC TTA CAG TG-3'                   Final concentration 100 ng/25µl

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 Unit/reaction
                                 (Perkin-Elmer Amplitaq DNA polymerase 5 U/µl, Cat. No. N808-0145)

PCR Cycling Profile: 95^oC (5')
                              95^oC (15"), 54^oC (15"), 72^oC (60")  × 40 cycles
                              72^oC (10')

Product size:
                                  StyI
                   477 bp ---------------> 263 bp + 214 bp (in homozygote for site present)
 
 

Kidd Lab Histograms for Allele Frequencies