Bacterial Cell Transformation and Genomic Libraries

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What is one of the reasons why vector DNA cannot re-ligate to itself?

Blunt ends of the digested vector DNA are incompatible (correct)

Excessive amount of DNA Ligase used

The size of the vector DNA is too small

Presence of cohesive ends on the digested vector DNA

Which components are required for making recombinant DNA molecules?

DNA Ligase, E.coli, Antibiotic selection

Plasmid vector, Insert DNA, Restriction enzyme (correct)

Insert DNA, DNA Ligase, Transformation

Insert DNA, E.coli, Restriction enzyme

Why is directional cloning made possible with insert DNA fragments?

They possess two different cohesive/sticky ends on either side (correct)

They lack cohesive ends

They have blunt ends

They are very large in size

Directional cloning allows the insert to be cloned in which specific orientation?

5' to 3' Signup and view all the answers

How can you determine if cloning has been successful in transformation?

By antibiotic selection Signup and view all the answers

What must be done to cells to allow them to take up DNA in the process of transformation?

Introducing a heat shock Signup and view all the answers

In constructing a cDNA library, what is the starting material used?

mRNA Signup and view all the answers

What is the function of genomic libraries?

Collecting individual clones representing the entire genome Signup and view all the answers

How do you verify that a plasmid is truly recombinant?

By observing if the inserted DNA fragment disrupts a gene within the plasmid Signup and view all the answers

What method is primarily used for generating transgenic bacteria?

Bacterial transformation by a recombinant plasmid Signup and view all the answers

To clone mRNA sequences into a vector, what type of cDNA copy must be synthesized first?

Double-stranded Signup and view all the answers

What is the main purpose of ligating cDNA into a vector in the construction of cDNA libraries?

Cloning vectors Signup and view all the answers

What is the purpose of using two different restriction enzymes in directional cloning?

To ensure the insert DNA is ligated into the vector in a specific orientation Signup and view all the answers

What are cohesive compatible ends in DNA cloning?

Complementary sticky ends created by two different restriction enzymes Signup and view all the answers

How does directional cloning ensure specific orientation of insert DNA in the vector?

By utilizing cohesive compatible ends Signup and view all the answers

What is one advantage of using directional cloning in DNA experiments?

Ensures all insert-DNA fragments are efficiently inserted into the vector Signup and view all the answers

In DNA cloning, what role do cohesive compatible ends play?

Ensure specific orientation of insert DNA in vectors Signup and view all the answers

What is the significance of using two restriction enzymes with different sequences in directional cloning?

Facilitates specific orientation of insert DNA in vectors Signup and view all the answers