This procedure will work for both yeast and E. coli: Take a small colony and suspend it in 5ul of H2O in a PCR tube. Heat for 5 min at 95℃ and then spin the condensation down in a microfuge. Set up the PCR reaction as follows: 5ul H2O + cells 5ul 5uM primer2 5ul 10×Taq Buffer 5ul 2mM dNTPs 0.5ul 10 mg/ml acetylated BSA 1ul Taq DNA polymerase 23.5 ul H2O PCR Conditions: 94℃×4min. then 35 cycles of: (94℃×1 min then 55℃× 1min then 72℃×3min) followed by 72℃×20 min and a 4℃ soak. (5ul run out on a mini gel should be sufficient to see product.) |
→如果您认为本词条还有待完善,请 编辑词条
上一篇Design of the Invader Genotyping Assay 下一篇PCR技术(七):mRNA差异PCR技术
词条内容仅供参考,如果您需要解决具体问题
(尤其在法律、医学等领域),建议您咨询相关领域专业人士。
0