Kidd Lab 

EcoRI site (located in intron 2)

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

Primer 1. A3EX2DW:    5'-TTG CAC CTC CTA AGG CTC-3'                   Final concentration 100 ng/25µl
Primer 2. A3EcoUP2:   5'-TCT AAT GCA AAT TGA TTG TGA AC-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
DMSO: Final Concentration 5%

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"), 51^oC (15"), 72^oC (75")  × 40 cycles
                              72^oC (10')

Product size:
                                 EcoRI
                   323 bp ---------------> 242 bp + 81 bp (in homozygote site present)
 
 

Kidd Lab Histograms for Allele Frequencies