Kidd Lab 

HaeIII site (located in exon 5)

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

Primer 1. A3EX5FOR2:    5'-TGA GTT TGC ACA TTA GTT ATG G-3'        Final concentration 100 ng/25µl
Primer 2. A3EX5REV1:    5'-TGC TCT CAG TTC TTT CTG GG-3'              Final concentration 100 ng/25µl

dNTPs: Final Concentration 200 µM/reaction

Buffer: Final Concentration 2.0 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 (30"), 56^oC (30"), 72^oC (60")  × 35 cycles
                             72^oC (10')

Product size:
                                 HaeIII
                   435 bp ---------------> 193 bp + 242 bp (in homozygote site present)
 
 

Kidd Lab Histograms for Allele Frequencies