A 3' RACE protocol to confirm polyadenylation sites

Liuyin Ma; Arthur G Hunt

doi:10.1007/978-1-4939-2175-1_12

A 3' RACE protocol to confirm polyadenylation sites

Methods Mol Biol. 2015:1255:135-44. doi: 10.1007/978-1-4939-2175-1_12.

Authors

Liuyin Ma¹, Arthur G Hunt

Affiliation

¹ College of the Environment and Ecology, Xiamen University, Xiamen, 361102, China.

PMID: 25487210
DOI: 10.1007/978-1-4939-2175-1_12

Abstract

A routine procedure in the study of polyadenylation involves the determinations of the junctions of the mRNA body and the poly(A) tail (the poly(A) site). This is typically accomplished by selectively amplifying the 3'-ends of cDNAs (3'-Rapid Amplification of cDNA Ends, or 3'-RACE), followed by sequencing of individual clones. We have developed a modification of the standard 3'-RACE protocol that couples high specificity with the ability to characterize thousands (or more) of individual sequences. This protocol may be used for numerous purposes, including the confirmation of results obtained using high throughput sequencing technologies.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

DNA, Complementary / chemistry*
Nucleic Acid Amplification Techniques
Poly A / chemistry
Poly A / genetics
Polyadenylation*
RNA, Messenger / chemistry*
RNA, Messenger / genetics
Sequence Analysis, DNA / methods

Substances

DNA, Complementary
RNA, Messenger
Poly A