en
Scientific article
English

Selective amplification via biotin- and restriction-mediated enrichment (SABRE), a novel selective amplification procedure for detection of differentially expressed mRNAs

Publication date1997
Abstract

We present a novel subtractive enrichment protocol for the identification of differentially expressed mRNA species. This procedure, called SABRE (selective amplification via biotin- and restriction-mediated enrichment), uses selective streptavidin-biotin affinity and restriction enzyme site reconstitution to enrich for cDNA species more abundant in one population than in another. Analysis of liver cDNA from a mouse strain expressing the neomycin resistance gene demonstrated that this procedure is capable of identifying species present in one population but absent from another. Furthermore, experiments to identify genes with circadian expression patterns in mouse liver demonstrated that SABRE is capable of detecting even modest 2- to 10-fold differences in accumulation of moderately rare mRNA species, representing as little as 0.03% of total mRNA. These experiments identified the gene encoding coumarin 7-hydroxylase as displaying circadian expression in mouse liver.

Keywords
  • Animals
  • Aryl Hydrocarbon Hydroxylases
  • Biotin
  • Circadian Rhythm
  • Cytochrome P-450 Enzyme System/genetics
  • DNA, Complementary/analysis/genetics
  • Humans
  • Liver/enzymology
  • Mice
  • Mixed Function Oxygenases/genetics
  • Nucleic Acid Hybridization
  • Polymerase Chain Reaction/methods
  • RNA, Messenger/analysis
Citation (ISO format)
LAVERY, Daniel et al. Selective amplification via biotin- and restriction-mediated enrichment (SABRE), a novel selective amplification procedure for detection of differentially expressed mRNAs. In: Proceedings of the National Academy of Sciences of the United States of America, 1997, vol. 94, n° 13, p. 6831–6836. doi: 10.1073/pnas.94.13.6831
Main files (1)
Article (Published version)
accessLevelRestricted
Identifiers
ISSN of the journal0027-8424
546views
0downloads

Technical informations

Creation12/09/2014 2:55:00 PM
First validation12/09/2014 2:55:00 PM
Update time03/14/2023 10:26:45 PM
Status update03/14/2023 10:26:45 PM
Last indexation01/16/2024 3:11:13 PM
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack