title: Measuring gene expression from cell cultures by quantitative reverse-transcriptase polymerase chain reaction.
abstract: Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) offers a robust method for the measurement of RNA levels for any gene within cells harvested at any point before or during cell culture. The key elements of RNA extraction followed by a two-step qRT-PCR method (reverse transcription and PCR) are described, followed by a brief section on analysis of the results. There are a number of excellent kits available commercially for much of this work, but it is essential to ensure that the quality and quantity of cDNA produced is adequate for the standard PCR or array to be used.
authors.affiliation: Translational Oncology Research Centre, Queen Alexandra Hospital, Portsmouth, UK.
authors.forename: Sharon
authors.initials: S
authors.lastname: Glaysher
authors.forename: Francis G
authors.initials: FG
authors.lastname: Gabriel
authors.forename: Ian A
authors.initials: IA
authors.lastname: Cree
mesh_terms: D002457:Cell Extracts; D002478:Cells, Cultured; D020869:Gene Expression Profiling / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D048348:Reverse Transcription; D019914:Taq Polymerase / Q000378:metabolism
chemical_list: D002457:Cell Extracts; D012313:RNA; D019914:Taq Polymerase