Description NGFR HincII Polymorphism Procedure HincII (PCR location in AC006487: 11430-12116 bp; HincII location in AC006487: 11499-11504bp 1695 bp downstream from Exon1 of NGFR) Template: Genomic DNA 100-200 ng/reaction volume of 25 µl. Primer 1. NGFRHNCF 5'-CAC CTT TTC AGG GGG CAG C-3' 100 ng/Reaction Primer 2. NGFRHNCR 5'-CCC TCG TTC CAT CCC TAT TA-3' 100 ng/Reaction dNTPs: Final Concentration 200 µM each Buffer: Final Concentration 1.15 mM MgCl2, 50 mM KCl, 10 mM Tris HCl pH 8.4, 1M betane NOTE THE BETANE Taq DNA Polymerase: 0.5 Units/reaction volume of 25 µl (Perkin Elmer Amplitaq DNA polymerase 5 U/µl, Cat. No. N808-0145) PCR Cycling Profile: 94ºC, 5 94ºC (1'), 58ºC (30"), 72ºC (30") × 10 cycles 94ºC (30"), 58ºC (30"), 72ºC (30") 25 cycles 72ºC (7'), Product size: HincII 687------------------> 615 + 72 bp (in +/+ homozygote) Recipes Supplies Tips (责任编辑:泉水) |