Abstract
Quantitation of mRNA by polymerase chain reaction requires valuation of the possible modification of native mRNA molecule through cDNA synthesis (see chapter 2.2), following amplification (see chapters 2.3 and 2.4) and quantitation of generated PCR products by several methods (see chapters 2.5; 3.1; 3.3; 3.4). Addition of an internal RNA standard during extraction step in RNA isolation protocol is the most reliable check of processing of the initial RNA molecule in complete RT-PCR. The use of DNA standards can also be successful (see also chapter 1.2). A disadvantage of using a DNA standard is the different transcription rate in cDNA synthesis.
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© 1995 Springer-Verlag Berlin Heidelberg
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Lassner, D. (1995). Cloning of Short DNA Fragments and In Vitro Transcription to Generate RNA Standards. In: Quantitation of mRNA by Polymerase Chain Reaction. Springer Labor Manuals. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-79712-5_3
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DOI: https://doi.org/10.1007/978-3-642-79712-5_3
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-79714-9
Online ISBN: 978-3-642-79712-5
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