In vitro transcription and immobilized template analysis of preinitiation complexes

In the study of gene regulation, it is often necessary to employ functional assays that investigate the action or mechanism of specific promoters or enhancer-binding factors and their role in transcription by RNA polymerase II. Although many assays measure the transcription of a gene under the contr...

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Bibliographic Details
Published inCurrent protocols in molecular biology (Print) Vol. Chapter 12; p. Unit 12.14.
Main Authors Lin, Justin J, Carey, Michael
Format Journal Article
LanguageEnglish
Published United States 01.01.2012
Subjects
Cell Nucleus - chemistry
Cell Nucleus - genetics
DNA - chemistry
DNA - genetics
DNA Primers - genetics
Genetic Techniques
Humans
RNA - genetics
Templates, Genetic
Transcription, Genetic
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Summary:In the study of gene regulation, it is often necessary to employ functional assays that investigate the action or mechanism of specific promoters or enhancer-binding factors and their role in transcription by RNA polymerase II. Although many assays measure the transcription of a gene under the control of an endogenous or model activator in vivo, it is often useful to recreate transcription in vitro in order to study specific regulatory mechanisms. In this unit, protocols are presented that will allow the investigator to perform in vitro transcription using preinitiation complexes assembled from cellular extracts on either naked DNA or chromatin templates.
ISSN:1934-3647
DOI:10.1002/0471142727.mb1214s97