Saturday, May 15, 2004
Description RET RsaI Polymorphism Procedure Rsa I Polymorphism in exon 15 Template: Genomic DNA 100-200 ng/reaction volume of 25 l. Primer 1. RET15RSA1 5'-GAC TCG TGC TAT TTT TCC TCA3' 100 ng/Reaction Primer 2. RET15RSA2 5'-TAT CTT TCC TAG GCT TCC CA 100 ng/Reaction dNTPs: Final Concentration 200 M each Buffer: Final Concentration 1.15 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"),562oC (30"), 72oC (30") 30 cycles Product size: RsaI 260 bp----------> 140 bp and 120 bp (in homozygote site present) Protocol from Ceccherini et al., 1994. Oncogene 9:3025-3029 Recipes Supplies Tips (责任编辑:泉水) |