Saturday, May 15, 2004
Description D10S94 MspI Polymorphism Procedure MspI Template: Genomic DNA 100-200 ng/reaction volume of 25 l Primer 1. S94MSPA 5 -CGA ATT CTT CCT GAA GCC T-3' 90 ng/reaction Primer 2. S94MSPB 5' -TTT GTA ATG GAG TGG TAT CT-3' 90 ng/reaction dNTPs: Final Concentration 200 M each Buffer: Final Concentration 1.5 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"), 54C (30"), 72C (30") x 30 cycles 72 C (7') Product size: Msp I 307 bp ----------->190 bp and 117 bp (in homozygote site present) Ref: Brooks-Wilson, A.R., D. Smailus, D. Gilchrist, and P.J. Goodfellow, 1992. Additional RFLPs at D10S94 and the development of PCR-based variant detection systems: Implications for disease genotype prediction in MEN2A, MEN2B, and MTC1 families. Genomics 13:233-234. Recipes Supplies Tips (责任编辑:泉水) |