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Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation
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Simultaneous control of DNA and RNA processing efficiency using a nucleic acid calibration set

Barbara Bartolini1, Anna Rosa Garbuglia, Douglas Horejsh

  • 1The Italian National Institute for Infectious Diseases, "L. Spallanzani"-IRCCS, Rome, Italy.

Biotechniques
|May 11, 2007
PubMed
Summary

No abstract available in PubMed .

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In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
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