Molecular Cloning Quiz 6

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Questions and Answers

What is the role of the recombinant plasmid in bacterial cells?

  • It helps in maintaining the sizes of DNA fragments.
  • It allows cells to express blue pigmentation.
  • It enables cells to become antibiotic resistant. (correct)
  • It prevents the cells from acquiring antibiotic resistance.

How can the orientation of an insert in a vector be tested?

  • By checking the antibiotic resistance of the bacteria.
  • By measuring the distance between the restriction sites.
  • By analyzing the color of bacterial colonies.
  • By using restriction enzyme C and observing fragment sizes. (correct)

Which statement is true regarding the color of bacterial cells that acquire a recombinant plasmid?

  • They are blue only if the insert is in the correct orientation. (correct)
  • They will become white if they do not acquire the plasmid.
  • They are always white regardless of the plasmid.
  • They turn blue when the vector contains DNA.

What is the purpose of ligation in molecular cloning?

<p>To integrate the insert into the vector. (D)</p> Signup and view all the answers

What does the compound Xgal do in cloning procedures?

<p>It interacts with the lacZ gene to produce a color change. (C)</p> Signup and view all the answers

What is the primary function of a restriction enzyme in molecular cloning?

<p>Cutting DNA at specific sequences (D)</p> Signup and view all the answers

Which of the following is typically used as a vector in bacterial cloning?

<p>Plasmid (A)</p> Signup and view all the answers

What is the process called where a foreign DNA fragment is inserted into a bacterium?

<p>Transformation (D)</p> Signup and view all the answers

What technique is used to identify bacterial colonies containing a specific construct using a selectable marker?

<p>Blue-white screening (C)</p> Signup and view all the answers

Which of the following enzymes is responsible for making a DNA copy from RNA?

<p>Reverse transcriptase (A)</p> Signup and view all the answers

Which of the following statements is false regarding recombinant plasmids?

<p>The cells with an empty vector are antibiotic resistant (B)</p> Signup and view all the answers

During blue-white screening, what indicates the presence of a recombinant plasmid?

<p>White colonies (D)</p> Signup and view all the answers

What is a key feature of plasmid replication?

<p>It requires host cell machinery but is independent of cell division (D)</p> Signup and view all the answers

Flashcards

Restriction Enzyme Function

Cuts DNA at specific sequences, crucial in molecular cloning.

Cloning Vector

A DNA molecule that carries foreign DNA into a host cell to replicate.

Bacterial Transformation

The process of inserting foreign DNA into a bacterium.

Blue-White Screening

A method to identify bacterial colonies containing a specific construct using a selectable marker.

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Reverse Transcriptase

Enzyme creating a DNA copy from RNA.

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Antibiotic Resistance

Cells with the recombinant plasmid are often antibiotic resistant.

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Plasmid Replication

Plasmid replication happens independently of the cell division.

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Xgal

A lactose analog that turns bacteria colonies blue when cleaved.

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Antibiotic resistance in bacterial cells

Some bacterial cells are not affected by antibiotics.

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Recombinant plasmid

A plasmid containing a new gene (or DNA) fragment.

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Molecular cloning steps

Preparing the gene of interest, the vector, ligation, transformation, and screening.

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Restriction enzyme choice

A specific restriction enzyme is chosen based on its cutting site in the gene's vector to ensure the correct orientation of insert.

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Restriction enzyme orientation test

Using a restriction enzyme to confirm the correct orientation of the inserted gene.

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Study Notes

True or False Questions (Molecular Cloning)

  • Cloning always selects blue colonies. False.
  • Colonies are blue due to disruption of the lacZ gene. True.
  • The gene of interest should always be in the lacZ gene. True.
  • Xgal produces blue coloration. True.
  • Restriction enzymes are used for both cloning and checking insert orientation. True.
  • The same restriction enzyme is used for both cloning steps. True.

Multiple Choice Questions (Molecular Cloning)

  • The main function of restriction enzymes is to cut DNA at specific sequences.
  • Plasmids are typically used as vectors in bacterial cloning.
  • Transformation is the process of inserting a foreign DNA fragment into bacterial cells.
  • Blue-white screening identifies bacterial colonies with the desired construct.
  • Reverse transcriptase creates a DNA copy from RNA.

Molecular Cloning Steps

  • Insert preparation
  • Vector preparation
  • Ligation
  • Transformation
  • Colony screening

Molecular Cloning Enzyme Selection

  • Enzyme B is preferred for cloning because it cuts upstream and downstream of the gene of interest but not within the gene itself.
  • Enzyme B also cuts a vector only once, within the lacZ gene

Molecular Cloning Restriction Enzyme Orientation Testing

  • Restriction enzyme C can be used to test the orientation of an insert.
  • If the insert is correctly oriented, digestion results in fragments of 230 bp and 5650 bp.
  • Incorrect orientation results in fragments of 650 bp and 5230 bp.

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Quiz 6: Molecular Cloning PDF

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