Journal
ANALYTICAL BIOCHEMISTRY
Volume 397, Issue 2, Pages 227-232Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2009.10.022
Keywords
5 ' RACE; PCR; Template-switching
Funding
- Swedish Energy Agency
- Knut and Alice Wallenberg Foundation
- Nordic Energy Research Program [BioH2]
- EU/NEST FP 6 [043340]
- EU/Energy FP 7 [212508]
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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.
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