Institut für Pharmazeutische Chemie, Philipps-Universität Marburg, Marburg, Germany.
Nucleic Acids Res. 2010 Aug;38(14):e147. doi: 10.1093/nar/gkq437. Epub 2010 May 26.
Here we describe a northern blot procedure that allows the detection of endogenous RNAs as small as approximately 14 nt in total RNA extracts from bacteria. RNAs that small and as part of total bacterial RNA extracts usually escape detection by northern blotting. The approach combines LNA probes 5'-digoxigenin-endlabeled for non-radioactive probe detection with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide-mediated chemical crosslinking of RNAs to nylon membranes, and necessitates the use of native PAGE either with the TBE or MOPS buffer system.
在这里，我们描述了一种 Northern blot 程序，该程序允许检测细菌总 RNA 提取物中大约 14 个核苷酸的内源性 RNA。如此小的 RNA 以及作为细菌总 RNA 提取物的一部分，通常会逃避 Northern blot 的检测。该方法结合了 LNA 探针 5'-地高辛标记，用于非放射性探针检测，以及 1-乙基-3-(3-二甲基氨基丙基）碳二亚胺介导的 RNA 与尼龙膜的化学交联，并且需要使用 TBE 或 MOPS 缓冲系统进行天然 PAGE。
Zotero 插件
