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Molecular Beacons and Fluorescence Detection
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Molecular Beacons and Fluorescence Detection

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Questions and Answers

Which structure does the probe form when the specific product is present?

  • Hairpin structure (correct)
  • Double helix structure
  • Single strand structure
  • Triple helix structure
  • When are Molecular Beacons detectable?

  • When the probes are bound to the template before displacement by the polymerase (correct)
  • When the probes are free in solution
  • When the probes are degraded
  • When the probes are bound to the template after displacement by the polymerase
  • What happens when the donor and acceptor probes are brought within 1 to 10 nm through specific DNA binding?

  • The acceptor loses its binding to the DNA
  • The donor loses its binding to the DNA
  • Excitation energy is transferred from the donor to the acceptor (correct)
  • The DNA structure changes
  • What is the role of the acceptor probe in Fluorescent Resonance Energy Transfer (FRET)?

    <p>The acceptor loses the energy in the form of heat or fluorescence emission</p> Signup and view all the answers

    Which factor is the amount of fluorescence directly proportional to?

    <p>The amount of template</p> Signup and view all the answers

    Which enzyme is responsible for cutting the FRET probe in the overlapping region?

    <p>Cleavase</p> Signup and view all the answers

    Which method of amplification uses a synthetic probe consisting of DNA and RNA sequences arranged in a DNA-RNA-DNA sandwich sequence?

    <p>Cycling Probe</p> Signup and view all the answers

    What is the role of the quencher dye in the cycling probe method of amplification?

    <p>To release the reporter dye</p> Signup and view all the answers

    How is the amount of target molecules in the sample related to the amount of signal produced in the FRET probe method?

    <p>The more target molecules, the more signal produced</p> Signup and view all the answers

    What is the purpose of the reporter molecule in the FRET probe method?

    <p>To produce signal</p> Signup and view all the answers

    Study Notes

    Molecular Beacons

    • In the presence of a specific product, the probe forms a stem-loop structure.

    Fluorescence Detection

    • Molecular Beacons are detectable when the specific product is present.

    FRET Basics

    • When the donor and acceptor probes are brought within 1 to 10 nm through specific DNA binding, energy transfer occurs from the donor to the acceptor.
    • The role of the acceptor probe in FRET is to absorb energy from the donor probe and emit it at a different wavelength.

    FRET Principles

    • The amount of fluorescence is directly proportional to the amount of target molecules in the sample.

    FRET Probe Cleavage

    • The enzyme responsible for cutting the FRET probe in the overlapping region is a nuclease.

    Cycling Probe Amplification

    • The cycling probe method of amplification uses a synthetic probe consisting of DNA and RNA sequences arranged in a DNA-RNA-DNA sandwich sequence.

    Quencher Dye Role

    • The quencher dye in the cycling probe method of amplification is responsible for absorbing the energy from the reporter molecule when the probe is intact.

    FRET Signal Production

    • The amount of signal produced in the FRET probe method is directly related to the amount of target molecules in the sample.

    Reporter Molecule Role

    • The purpose of the reporter molecule in the FRET probe method is to emit fluorescence when the probe is cleaved, indicating the presence of the target molecule.

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    Description

    Test your knowledge on Molecular Beacons and their fluorescence detection in this quiz. Learn about the factors affecting the amount of fluorescence and the relationship with the template. Find out if the fluorescence is directly or inversely proportional to the template amount.

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