RNA Detection and Quantification qRT-PCR
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Questions and Answers

What is a disadvantage of Northern blotting compared to Digital RNA counting?

  • Provides information on RNA length
  • Requires radioactive labelling (correct)
  • Works by using two probes
  • Can measure up to 800 targets

    • Why is formaldehyde mentioned in relation to RNA in the text?

  • To highlight its role as a primer in Digital RNA counting
  • To describe its function in cDNA synthesis
  • To explain its unfolding effect in Northern blotting (correct)
  • To emphasize its use in PCR amplification

    • What is a key advantage of Digital RNA counting over Northern blotting?

  • Can measure RNA length
  • Involves incubation at 42°C
  • Requires radioactive labelling
  • Provides a high throughput of up to 800 targets (correct)

    • Why might Digital RNA counting be considered expensive for small experiments?

    <p>Needs a big facility</p> Signup and view all the answers

    In Digital RNA counting, what is the function of the capture probe?

    <p>Tether the molecule to a glass plate</p> Signup and view all the answers

    What is the main limitation of Northern blotting in terms of throughput?

    <p>Can only measure a few RNA species at a time</p> Signup and view all the answers

    What is the primary purpose of qRT-PCR?

    <p>To quantify and detect RNA molecules</p> Signup and view all the answers

    What is the main limitation of qRT-PCR mentioned in the text?

    <p>It assumes equal cDNA synthesis efficiency across genes</p> Signup and view all the answers

    What technique can be used to overcome the limitation of qRT-PCR regarding RNA molecule size?

    <p>Northern blotting</p> Signup and view all the answers

    What is the first step in the Northern blotting technique?

    <p>RNA extraction and denaturation</p> Signup and view all the answers

    What is the purpose of pre-hybridization buffer in Northern blotting?

    <p>To reduce non-specific binding</p> Signup and view all the answers

    What is the final step in the Northern blotting technique described in the text?

    <p>Incubation in pre-hybridization buffer</p> Signup and view all the answers

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