Immuno-Northern Blotting: Detection of Modified RNA Using Gel Separation and Antibodies to Modified Nucleosides

免疫印迹法:利用凝胶分离和修饰核苷抗体检测修饰 RNA

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作者:Eikan Mishima, Takaaki Abe

Abstract

Immuno-northern blotting is a method for detecting modified RNAs using gel separation and specific antibodies to modified nucleosides. This method was developed by combining two commonly used molecular biology techniques: western blotting and northern blotting. In this method, urea-polyacrylamide (or agarose) gel-separated RNAs are transferred to positively charged nylon membrane and then immune detection is performed with specific antibodies to modified nucleosides: such as 1-methyladenosine, N6-methyladenosine, and pseudouridine. This highly sensitive and relatively simple method, which uses widely available laboratory equipment, enables small laboratories to compare the abundance of modified nucleic acids across samples.

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