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 Amplifi cation of cDNA Ends, or 3′-RACE), followed by sequencing of individual clones. We have developed a modifi cation of the standard 3′-RACE protocol that couples high specifi city with the ability to characterize thousands (or more) of individual sequences. This protocol may be used for numerous purposes, including the confi rmation of results obtained using high throughput sequencing technologies.
|Number of pages||10|
|Journal||Methods in Molecular Biology|
|State||Published - 2015|
Bibliographical notePublisher Copyright:
© Springer Science+Business Media New York 2015.
- 3′ RACE protocol
- Ion torrent
- Polyadenylation sites
ASJC Scopus subject areas
- Molecular Biology