Protocol

Analysis of Pre-mRNA Splicing Using HeLa Cell Nuclear Extracts

Adapted from RNA: A Laboratory Manual, by Donald C. Rio, Manuel Ares Jr, Gregory J. Hannon, and Timothy W. Nilsen. CSHL Press, Cold Spring Harbor, NY, USA, 2011.

Abstract

This protocol is used to determine the splicing behavior of pre-mRNAs in cell extracts that are capable of carrying out splicing (e.g., nuclear extracts from HeLa cells). 32P-labeled RNA is incubated under splicing conditions for various times, and the resulting products are analyzed on denaturing polyacrylamide gels.

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