title: Accurate RT-qPCR gene expression analysis on cell culture lysates.
abstract: Gene expression quantification on cultured cells using the reverse transcription quantitative polymerase chain reaction (RT-qPCR) typically involves an RNA purification step that limits sample processing throughput and precludes parallel analysis of large numbers of samples. An approach in which cDNA synthesis is carried out on crude cell lysates instead of on purified RNA samples can offer a fast and straightforward alternative. Here, we evaluate such an approach, benchmarking Ambion’s Cells-to-CT kit with the classic workflow of RNA purification and cDNA synthesis, and demonstrate its good accuracy and superior sensitivity.
authors.affiliation: Center for Medical Genetics, Ghent University Hospital , 9000 Ghent, Belgium.
authors.forename: Gert
authors.initials: G
authors.lastname: Van Peer
authors.forename: Pieter
authors.initials: P
authors.lastname: Mestdagh
authors.forename: Jo
authors.initials: J
authors.lastname: Vandesompele