Kidd Lab 

TaqI site (located in intron 7)

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

Primer 1.     ACADMTAQI:    5'-GAT CCA GTG ACT CAG AAG-3'                  100 ng/reaction
Primer 2.     ACADMTAQ2:   5'-TGA TAT TTC GAA AGA CAA AGA G-3'        100 ng/reaction

dNTPs: Final Concentration 200 µM each

Buffer: Final Concentration 2.25 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: 95^oC (5')
                              95^oC (30"), 56^oC (30"), 72^oC (30")  × 30 cycles

Product size:
                                  TaqI
                   162 bp  -----------> 113 bp + 49 bp (in homozygote site present)

Modified from Zhang, Z., S. Kolvraa, Y. Zhou, D.P. Kelly, N. Gregersen, and A.W. Strauss, 1993.  Three RFLPs defining a haplotype associated with the common mutation in human Medium-Chain Acyl-CoA Dehydrogenase (MCAD) deficiency occur in Alu repeats. Am. J. Hum. Genet. 52: 1111-1121.
 
 

Kidd Lab Histograms for Allele Frequencies