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Semi-quantitative RT-PCR for comparison of mRNAs in cells with different amounts of housekeeping gene transcripts

https://doi.org/10.1006/mcpr.1998.0182Get rights and content

Abstract

A simple method is reported here for the semi-quantitative assay of mRNAs in the presence of an exogenous mRNA (pBR322 transcript) as a standard. This method uses the co-reverse transcription and co-amplification (co RT-PCR) of the target and standard mRNAs. This procedure enables transcripts to be compared when the differentiation process affects the transcription pattern of the β-actin housekeeping gene, a commonly used internal standard. This method is sensitive and avoids constructing internal competitive RNA standards. As an example, it is shown that β-actin transcription decreases and type V adenylyl cyclase transcription increases in adipocytes, when compared to preadipocytes.

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Author to whom all correspondence should be addressed at: Laboratoire de Biochimie, Hôpital R Poincaré, F92380 Garches France.

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