Kidd Lab 

PstI site

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

Primer 1. CYP2EJ8:     5'-TTC ATT CTG TCT TCT AAC TGG-3'        final concentration 0.4µM
Primer 2. CYP2EJ9:     5'-CCA GTC GAG TCT ACA TTG TCA-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:
                                      PstI
                        410 bp--------------> 290 bp + 120 bp (in homozygote for site present)

Reference:
Shin-ichi Hayashi, et al., J. Biochem. 110: 559-565 (1991)
Fumika Tanaka, et al., Alcoholism: Clinical & Experimental Res.21, No. 4, (June 1997)
 
 

Kidd Lab Histograms for Allele Frequencies