High-throughput real-time PCR for detection of gene-expression levels.
While many high-throughput screening campaigns involve the measurement of protein levels or locations, at times it is desirable to measure the levels of gene expression in response to small molecules. Here, we describe a method for capturing mRNA in multiwell plates following compound treatment and measuring gene expression using real-time PCR. This streamlined protocol provides complementary information to conventional phenotypic cell-based assays, and is especially useful in cases where the gene of interest is thought to serve a regulatory function in downstream cellular phenotypes.
Chemical Biology Program, Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA. bwagner@broad.harvard.edu
Bridget K
BK
Wagner
Zoltan
Z
Arany
D000818:Animals; D002454:Cell Differentiation; D002477:Cells / Q000187:drug effects*; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015870:Gene Expression / Q000187:drug effects*; D051379:Mice; D009132:Muscles / Q000166:cytology / Q000187:drug effects; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*
D012333:RNA, Messenger