| 查看: 2164 | 回复: 1 | |||
[求助]
RNA与镁离子
|
|
求助下面这段描述中的镁离子是有什么作用的？另外镁离子与RNA的稳定与构象有关吗？原理是什么呢？ A T7 primer and gene-specific primers were used to PCR amplify pri-miR- 1792 sequences from plasmid DNA template. PCR products were gelpurified and used as templates for in vitro transcription using the Riboprobe (Promega) system together with 32P-CTP for radioactive labeling. Microprocessor purified from Flag-DROSHA-293 cells was used for in vitro Microprocessor assays (Gregory et al., 2004). Cold RNA was produced using the same strategy without 32P-CTP addition. For RNA annealing, 10 mM Sample Text was added to 200 pmol cold RNA and incubated at 95C for 5 min and then slowly cooled to reverse transcription (RT). Annealed RNA was subjected to 5% native polyacrylamide gel for ethidium bromide staining and used for Microprocessor assay followed by small RNA northern blot analysis. His- CPSF3 complex was purified from E. coli as described previously for other proteins (Chang et al., 2013). Assays conditions were as for Microprocessor. |
回复此楼» 猜你喜欢
湖北师范大学招调剂啦
已经有9人回复
-
邵阳学院食品与化学工程学院硕士调剂
已经有0人回复
-
化学工程及工业化学论文润色/翻译怎么收费?
已经有260人回复
-
天津商业大学 生物与医药课题组招生
已经有0人回复
-
生物化工学硕招收调剂
已经有0人回复
-
广东石油化工学院2026年硕士研究生需要多名生物，制药工程，食品专业的调剂生
已经有0人回复
-
化工申博
已经有3人回复
-
申博自荐
已经有3人回复
-
广西民族大学化学化工学院化学工程与技术学硕，材料化工专硕调剂名额充足！
已经有0人回复
|
In Vitro Transcription, Microprocessor, and CPSF3 Cleavage Assays A T7 primer and gene-specific primers were used to PCR amplify pri-miR- 1792 sequences from plasmid DNA template. PCR products were gelpurified and used as templates for in vitro transcription using the Riboprobe (Promega) system together with 32P-CTP for radioactive labeling. Microprocessor purified from Flag-DROSHA-293 cells was used for in vitro Microprocessor assays (Gregory et al., 2004). Cold RNA was produced using the same strategy without 32P-CTP addition. For RNA annealing, 10 mM MgCl2 was added to 200 pmol cold RNA and incubated at 95C for 5 min and then slowly cooled to reverse transcription (RT). Annealed RNA was subjected to 5% native polyacrylamide gel for ethidium bromide staining and used for Microprocessor assay followed by small RNA northern blot analysis. His- CPSF3 complex was purified from E. coli as described previously for other proteins (Chang et al., 2013). Assays conditions were as for Microprocessor. |
2楼2015-09-23 18:37:14