Gibson Assembly for Long Inserts
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Questions and Answers

What type of ends does an insert with flanking Bsa1 sites have?

  • Blunt ends
  • Circular ends
  • Sticky ends (correct)
  • Flared ends
  • Which process is used to join multiple sequences like a train in Golden Gate Cloning?

  • Polymerization
  • Ligation (correct)
  • Digestion
  • Annealing
  • What method is used for RNA quantification and detection?

  • Gel electrophoresis
  • Mass spectrometry
  • Western blot
  • qRT-PCR (correct)
  • What are the Bsa1 sites used for in vector construction?

    <p>To remove recognition sites</p> Signup and view all the answers

    What enzyme is essential for the quantitative PCR process?

    <p>DNA polymerase</p> Signup and view all the answers

    In Golden Gate Cloning, what is the function of divergent Bsa1 sites in the destination vector?

    <p>Flank the selection marker gene</p> Signup and view all the answers

    Which technique is used to clone long inserts by assembling multiple DNA fragments together?

    <p>Gibson assembly</p> Signup and view all the answers

    Which enzymes are used in the Gibson assembly method?

    <p>5' exonuclease, DNA polymerase, and DNA ligase</p> Signup and view all the answers

    What is the potential issue with repeating sequences in the DNA fragments used for Gibson assembly?

    <p>The fragments may anneal at the wrong places</p> Signup and view all the answers

    Which cloning method allows for the assembly of multiple sequence inserts without generating a single stretch of single-stranded DNA?

    <p>Golden Gate cloning</p> Signup and view all the answers

    What is the unique property of the Type IIS restriction enzymes used in Golden Gate cloning?

    <p>They cleave at a specific distance from their recognition sequence</p> Signup and view all the answers

    Which of the following statements about the Type IIS restriction enzymes used in Golden Gate cloning is true?

    <p>They cleave DNA in a directional manner</p> Signup and view all the answers

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