Saturday, May 15, 2004
Description RET TaqI Polymorphism Procedure Taq I polymorphism located in Exon 13 Template: Genomic DNA 50-100 ng/reaction volume of 25 ul Primer 1. RETEX13F: 5'-CTC TCT GTC TGA ACT TGG GC-3' 60 ng/Reaction Primer 2. RETEX13R: 5'-TCA CCC TGC AGC AGG CCT TA-3' 60 ng/Reaction dNTPs: Final Concentration in the reaction 333 uM each Buffer: Final Concentration 2.25 mM MgCl2, 50 mM KCl, 10 mM Tris HCl pH 8.4 BSA: Final Concentration 16 ug/ul (e.g., .5 l @ 10 mg/ml BSA/30 ul Rx) DMSO: Final Concentration 5% Taq DNA Polymerase: 0.75 U/reaction (Perkin Elmer Amplitaq DNA polymerase 5 U/ul, Cat. No. N808-0145) PCR Cycling Profile: 95oC, 5' 95oC (30"), 56oC (30"), 72oC (1') x 30 Product Size: TaqI 239 -----------> 133 + 106 (in homozygote site present) Romeo, G., P. Ronchetto, Y. Luo, V. Barone, M. Seri, I. Ceccherini, B. Pasini, R. Bocciardi, M. Lerone, H. Kaariainen, et al., 1994. Point mutations affecting the tyrosine kinase domain of the RET proto-oncogene in Hirschsprung's disease. Nature 367:377-378. Recipes Supplies Tips (责任编辑:泉水) |