title: Absolute and relative gene expression in Listeria monocytogenes using real-time PCR.
abstract: Reverse transcription combined with the polymerase chain reaction (RT-PCR) is a viable method widely used to quantify gene expression. There are two ways to quantify gene expression by real-time PCR: relative quantification and absolute quantification. Relative quantification relates the PCR signal of the target gene to a control gene, normally a housekeeping gene. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Here we describe both methods from RNA extraction to its quantification by real-time PCR.
authors.affiliation: Department of Veterinary Medicine, University of Sassari, Via Vienna 2, 07100, Sassari, Italy, rmazza@uniss.it.
authors.forename: Roberta
authors.initials: R
authors.lastname: Mazza
authors.forename: Rina
authors.initials: R
authors.lastname: Mazzette
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors / Q000235:genetics; D008089:Listeria monocytogenes / Q000235:genetics*; D008088:Listeriosis / Q000382:microbiology; D010957:Plasmids / Q000235:genetics / Q000302:isolation & purification; D012329:RNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D018076:DNA, Complementary; D012329:RNA, Bacterial