Kidd Lab 

AluI site (located in exon 7)

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

Primer 1.     PAHALU333:    5'-CTT GCA CTG GTT TCC GCC TC-3'              100 ng/reaction
Primer 2.     PAHALU237:    5'-CCC AAA CCT CAT TCT TGC AGC A-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
                                 (Perkin-Elmer Amplitaq DNA polymerase 5 U/µl, Cat. No. N808-0145)

PCR Cycling Profile:  95^oC (5')
                               95^oC (30"), 59^oC (30"), 72^oC (30")  × 30 cycles

Product size:
                               AluI
                  213 bp ------------> 124 bp +  59 bp + 30 bp (in AluI site present homozygote)
                  213 bp ------------> 154 bp +  59 bp (in AluI site-absent homozygote)
                  NOTE: the 59 bp fragment is a constant fragment.
 
 

Kidd Lab Histograms for Allele Frequencies