Kidd Lab 

MwoI site (located in intron 2)

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

Primer 1.     ADH6MWOF:          5'-CTG TCT GGC TAA CTT ACA ATA AAG-3'        100 ng/reaction
Primer 2.     ADH6INT2B:           5'-TGT GGA GGA GAA AGA ACG ACT-3'             100 ng/reaction

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: 0.5 Units/reaction volume of 25 µl
                                 (Perkin-Elmer Amplitaq DNA polymerase 5 U/µl, Cat. No. N808-0145)

PCR Cycling Profile: Use a Touch Down protocl:
                               94^oC(5')
                              95^oC (30"), 65^oC (30"), 72^oC (45")  × 3 cycles
                              95^oC (30"), 63^oC (30"), 72^oC (45")  × 3 cycles
                              95^oC (30"), 61^oC (30"), 72^oC (45")  × 32 cycles
                              72^oC (7')

Product size:
                                MwoI
                    483 bp-------------> 261 bp + 222 bp (in homozygote site present)
 
 

Kidd Lab Histograms for Allele Frequencies