18 Questions
What is one of the reasons why vector DNA cannot re-ligate to itself?
Blunt ends of the digested vector DNA are incompatible
Which components are required for making recombinant DNA molecules?
Plasmid vector, Insert DNA, Restriction enzyme
Why is directional cloning made possible with insert DNA fragments?
They possess two different cohesive/sticky ends on either side
Directional cloning allows the insert to be cloned in which specific orientation?
5' to 3'
How can you determine if cloning has been successful in transformation?
By antibiotic selection
What must be done to cells to allow them to take up DNA in the process of transformation?
Introducing a heat shock
In constructing a cDNA library, what is the starting material used?
mRNA
What is the function of genomic libraries?
Collecting individual clones representing the entire genome
How do you verify that a plasmid is truly recombinant?
By observing if the inserted DNA fragment disrupts a gene within the plasmid
What method is primarily used for generating transgenic bacteria?
Bacterial transformation by a recombinant plasmid
To clone mRNA sequences into a vector, what type of cDNA copy must be synthesized first?
Double-stranded
What is the main purpose of ligating cDNA into a vector in the construction of cDNA libraries?
Cloning vectors
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
What are cohesive compatible ends in DNA cloning?
Complementary sticky ends created by two different restriction enzymes
How does directional cloning ensure specific orientation of insert DNA in the vector?
By utilizing cohesive compatible ends
What is one advantage of using directional cloning in DNA experiments?
Ensures all insert-DNA fragments are efficiently inserted into the vector
In DNA cloning, what role do cohesive compatible ends play?
Ensure specific orientation of insert DNA in vectors
What is the significance of using two restriction enzymes with different sequences in directional cloning?
Facilitates specific orientation of insert DNA in vectors
Learn about the process of transforming bacterial cells by making them competent to take up DNA, including the steps of dilute salt solution, heat shock, and subsequent DNA replication and expression of antibiotic resistance. Explore the construction of genomic libraries from genomic DNA of organisms.
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