Selective amplification via biotin- and restriction-mediated enrichment (SABRE), a novel selective amplification procedure for detection of differentially expressed mRNAs
| Autor: | Ueli Schibler, Daniel J. Lavery, Fabienne Fleury-Olela, Luis Lopez-Molina |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1997 |
| Předmět: |
DNA
Complementary Polymerase Chain Reaction/methods Population Cytochrome P-450 Enzyme System/genetics Biotin Biology Polymerase Chain Reaction Mixed Function Oxygenases law.invention Cytochrome P-450 CYP2A6 Nucleic acid thermodynamics chemistry.chemical_compound Mice Cytochrome P-450 Enzyme System law Complementary DNA ddc:570 Animals Humans RNA Messenger Circadian rhythm education Gene Polymerase chain reaction education.field_of_study Messenger RNA Multidisciplinary RNA Messenger/analysis Liver/enzymology Nucleic Acid Hybridization Biological Sciences Molecular biology Circadian Rhythm ddc:580 DNA Complementary/analysis/genetics Liver chemistry Mixed Function Oxygenases/genetics Aryl Hydrocarbon Hydroxylases |
| Zdroj: | Proceedings of the National Academy of Sciences, Vol. 94, No 13 (1997) pp. 6831-6836 |
| ISSN: | 0027-8424 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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