Saturday, May 15, 2004
Description PAH AluI Polymorphism Procedure AluI; LOCATED IN EXON 7 Template: Genomic DNA 100-200 ng/reaction volume of 25 l Primer 1. PAHALU333 5' CTTGCACTGGTTTCCGCCTC 3' 100 ng/reaction Primer 2. PAHALU237 5' CCCAAACCTCATTCTTGCAGCA 3' 100 ng/reaction dNTPs: Final Concentration 200 M each Buffer: Final Concentration 1.50 mM MgC12, 50 mM KCl, 10 mM Tris HCl pH 8.4 Taq DNA Polymerase: 0.5 Units/reaction (Perkin-Elmer Amplitaq DNA polymerase 5 U/l, Cat. No. N808-0145) PCR Cycling Profile: 95C, 5' 95C (30"), 59C (30"), 72C (30") x 30 cycles Product size: AluI 213 bp -----------> 124, 59 and 30 bp in AluI site present homozygote; 213 bp------------> 154 and 59 bp in AluI site-absent homozygote NOTE: the 59 bp fragment is a constant fragment Recipes Supplies Tips (责任编辑:泉水) |