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

Sammanfattning

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.

Nyckelord

5 ' RACE; PCR; Template-switching

Publicerad i

Analytical Biochemistry
2010, volym: 397, nummer: 2, sidor: 227-232
Utgivare: ACADEMIC PRESS INC ELSEVIER SCIENCE

SLU författare

• Lopes Pinto, Fernando A

  • Uppsala universitet

UKÄ forskningsämne

Biokemi
Molekylärbiologi

Publikationens identifierare

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

Permanent länk till denna sida (URI)

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