title: The Use of Real-Time Reverse Transcription-PCR for Assessing Estrogen Receptor and Estrogen-Responsive Gene Expression.
abstract: Real-time reverse transcription-polymerase chain reaction (RT-PCR), also known as quantitative RT-PCR (qRT-PCR), is a powerful tool for assessing gene transcription levels. The technique is especially useful for measuring estrogen receptor transcript levels as well as gene expression changes in response to estrogen stimulation as it is quick, accurate, robust, and allows the measurement of gene expression in a variety of tissues and cells. This chapter describes the protocols used for the real-time RT-PCR assay using hydrolysis (TaqMan-type) probes.
authors.affiliation: Sanford Research, 2301 East 60th Street North, Sioux Falls, SD, 57104, USA. Michelle.Booze@SanfordHealth.org.
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Booze
authors.affiliation: Division of Basic Biomedical Sciences, Sanford School of Medicine, University of South Dakota, 414 E. Clark St., Vermillion, SD, 57069, USA.
authors.forename: Kathleen M
authors.initials: KM
authors.lastname: Eyster
mesh_terms: D000818:Animals; D002460:Cell Line; D015342:DNA Probes; D004967:Estrogens / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D006868:Hydrolysis; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D011960:Receptors, Estrogen / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019914:Taq Polymerase / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D057188:Workflow
chemical_list: D015342:DNA Probes; D004967:Estrogens; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D019914:Taq Polymerase
keywords: Custom design; Estrogen receptor s; Fluorogenic probe; Gene expression; Master mix; Normalization; Primer; Real-time RT-PCR