Control selection for RNA quantitation

The study of mammalian gene expression is often carried out at the level of mRNA. In such analyses, one usually measures the amount of an mRNA of interest under different conditions such as stress, growth, development, cell and tissue localization or as part of an evaluation of the effects of gene t...

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Détails bibliographiques
Publié dans:BioTechniques. - 1991. - 29(2000), 2 vom: 08. Aug., Seite 332-7
Auteur principal: Suzuki, T (Auteur)
Autres auteurs: Higgins, P J, Crawford, D R
Format: Article
Langue:English
Publié: 2000
Accès à la collection:BioTechniques
Sujets:Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S. Review Validation Study Actins Cytokines DNA, Ribosomal Heat-Shock Proteins Hormones plus... RNA, Messenger RNA, Ribosomal Tubulin Manganese 42Z2K6ZL8P Glyceraldehyde-3-Phosphate Dehydrogenases EC 1.2.1.- Calcitriol FXC9231JVH
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520 |a The study of mammalian gene expression is often carried out at the level of mRNA. In such analyses, one usually measures the amount of an mRNA of interest under different conditions such as stress, growth, development, cell and tissue localization or as part of an evaluation of the effects of gene transfection. A variety of techniques exist to measure gene expression and most commonly involve Northern hybridization analysis, ribonuclease protection or RT-PCR. Common to all of these assays is the inclusion of a so-called loading or internal control (i.e., analysis of an mRNA that does not change in relative abundance during the course of treatments). Here, we discuss the uses and pitfalls of the most popular of these controls, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and beta-actin, with special emphasis on precautions associated with the use of GAPDH 
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