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Reverse-Transcriptase Polymerase Chain Reaction to Detect Extracellular mRNAs

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Gene Expression Profiling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 784))

Abstract

The presence of extracellular nucleic acids has been reported in serum/plasma from cancer and diabetes patients that may help in disease diagnosis. Taking insulin-producing cells as examples here, RT-PCR was used to investigate a correlation between the presence and amounts of extracellular mRNA(s) and cell mass and/or function. RT-PCR was performed on a range of mRNAs, including Pdx1, Npy, Egr1, Pld1, Chgb, InsI, InsII, and Actb in biological triplicate analyses.

Reproducible amplification of these mRNAs from MIN6, MIN6 B1, and Vero-PPI cells and their CM suggests that beta cells transcribe and release these mRNAs into their environment. mRNAs secreted from insulin-producing cells into their extracellular environment may have potential as extracellular biomarkers for assessing beta cell mass and function.

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Correspondence to Sweta Rani .

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Rani, S., O’Driscoll, L. (2011). Reverse-Transcriptase Polymerase Chain Reaction to Detect Extracellular mRNAs. In: O'Driscoll, L. (eds) Gene Expression Profiling. Methods in Molecular Biology, vol 784. Humana Press. https://doi.org/10.1007/978-1-61779-289-2_2

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  • DOI: https://doi.org/10.1007/978-1-61779-289-2_2

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-288-5

  • Online ISBN: 978-1-61779-289-2

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