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. Recipes Supplies Tips (责任编辑:泉水) |