Saturday, May 15, 2004

Description
ACADM TaqI Polymorphism

Procedure
TaqI site (located in intron 7)

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

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 M 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 l
(Perkin-Elmer Amplitaq DNA polymerase 5 U/l, Cat. No. N808-0145)

PCR Cycling Profile: 95oC (5')
95oC (30"), 56oC (30"), 72oC (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.


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