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Control Selection for RNA Quantitation
Open accessAbstract
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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Cited by
Vandesompele, J., De Preter, K., Pattyn, F., Poppe, B., Van Roy, N., De Paepe, A., & Speleman, F. (2002) · Genome Biology · doi:10.1186/gb-2002-3-7-research0034 · Cited by 20,038
Andersen, C. L., Jensen, J. L., & Ørntoft, T. F. (2004) · Cancer Research · doi:10.1158/0008-5472.can-04-0496 · Cited by 7,127
Pfaffl, M. W., Tichopad, A., Prgomet, C., & Neuvians, T. P. (2004) · Biotechnology Letters · doi:10.1023/b:bile.0000019559.84305.47 · Cited by 5,065
Guideline to reference gene selection for quantitative real-time PCR
Radonić, A., Thulke, S., Mackay, I. M., Landt, O., Siegert, W., & Nitsche, A. (2004) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2003.11.177 · Cited by 1,643
Twenty-Five Years of Quantitative PCR for Gene Expression Analysis
VanGuilder, H. D., Vrana, K. E., & Freeman, W. M. (2008) · BioTechniques · doi:10.2144/000112776 · Cited by 1,246
Metadata
- Authors
- Suzuki, T., Higgins, P. J., & Crawford, D. R.
- Venue
- BioTechniques
- Publisher
- Informa UK Limited
- Published
- 2000-08
- Volume
- 29
- Issue
- 2
- Pages
- 332-337
- ISSN
- 0736-6205 (ISSN-L)
- 1940-9818
- OpenAlex
- W2131987407
- Cited by
- 875
- References
- 0
- FWCI
- 7.66
- Type
- journal article
- Language
- en
- Open access
- gold
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