Kidd Lab 

MspI site (located in intron 6)

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

Primer 1.     CYP2E3:     5'-CTG TGC TGA ACA CTG GTA CTC-3'        final concentration 0.4µM
Primer 2.     CYP2E6:     5'-ACT GGC CTG TTT GCA TGC G-3'            final concentration 0.4µM

dNTPs: Final Concentration 200 µM each

Buffer: Final Concentration 1.5 mM MgCl₂, 50 mM KCl, 10 mM Tris-HCl pH 8.4

Taq DNA Polymerase: 1 Unit/reaction
                                 (Perkin-Elmer Amplitaq DNA polymerase 5 U/µl, Cat. No. N808-0145)

PCR Cycling Profile: 94^oC (5')
                              94^oC (40"), 58^oC (30"), 72^oC (1')  × 30 cycles
                              72^oC (10')

Product size:
                                 MspI
                    480 bp--------------> 330 bp + 150 bp (in homozygote for site present)
 

Kidd Lab Histograms for Allele Frequencies