1. A guide for in-house design of template-switch-based 5' rapid amplification of cDNA ends systems
- Author
-
Fernando Lopes Pinto and Peter Lindblad
- Subjects
0106 biological sciences ,Untranslated region ,DNA, Complementary ,Biophysics ,Biology ,01 natural sciences ,Biochemistry ,Polymerase Chain Reaction ,03 medical and health sciences ,Rapid amplification of cDNA ends ,RNA, Messenger ,Molecular Biology ,030304 developmental biology ,Genetics ,0303 health sciences ,Messenger RNA ,Transcription start ,Reproducibility of Results ,RNA-Directed DNA Polymerase ,Cell Biology ,Templates, Genetic ,Leukemia Virus, Murine ,Transcription Initiation Site ,5' Untranslated Regions ,Applications of PCR ,Nucleic Acid Amplification Techniques ,010606 plant biology & botany - 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.
- Published
- 2009