A guide for in-house design of template-switch-based 5 ' rapid amplification of cDNA ends systems

Abstract

Rapid amplification of cDNA ends (RACE) is an established strategy used to determine the transcription start point(s) and the 5' untranslated region(s) of mRNA. Different approaches to perform 5' RACE are available, and one particularly simple and powerful strategy is based on a phenomenon called template-switching. We investigated different aspects of template-switch-based 5' RACE, and we describe the different steps leading to the in-house development of a complete 5' RACE system-from oligonucleotide design to polymerase chain reaction (PCR) amplification. We show that the resulting system is reliable, time-efficient, and inexpensive. (C) 2009 Elsevier Inc. All rights reserved.

Keywords

5 ' RACE; PCR; Template-switching

Published in

Analytical Biochemistry
2010, Volume: 397, number: 2, pages: 227-232 Publisher: ACADEMIC PRESS INC ELSEVIER SCIENCE

UKÄ Subject classification

Biochemistry and Molecular Biology


Publication identifier

DOI: https://doi.org/10.1016/j.ab.2009.10.022

Permanent link to this page (URI)

https://res.slu.se/id/publ/87787