Question

Which of the following statements best describes how most sequences are inserted into cloning vectors?

(A) Both the insert and the cloning vectors are digested with restriction enzymes, providing Blunt Ends, and then the two are ligated together.
(B) Both the insert and the cloning vectors are digested with restriction enzymes, providing Blunt Ends. To prevent those cleaved sequences from recombining with their smaller fragments, the restriction digest products undergo Gel Electrophoresis, and the largest band is extracted from the gel. Then, the insert and vector are ligated together.
(C) Both the insert and the cloning vectors are digested with restriction enzymes, providing Sticky Ends, and then the two are ligated together.
(D) Both the insert and the cloning vectors are digested with restriction enzymes, providing Sticky Ends. To prevent those cleaved sequences from recombining with their smaller fragments, the restriction digest products undergo Gel Electrophoresis, and the largest band is extracted from the gel. Then, the insert and vector are ligated together.

Answer 1

The most accurate description of the process for inserting sequences into cloning vectors involves using restriction enzymes to create sticky ends on both the vector and the insert, after which they are ligated together.

Step-by-step explanation:

The question asks which of the following statements best describes how most sequences are inserted into cloning vectors. The best description is (C) Both the insert and the cloning vectors are digested with restriction enzymes, providing Sticky Ends, and then the two are ligated together. This process begins with the digestion of both the cloning vector and the DNA insert with restriction enzymes that create complementary sticky ends. These ends can anneal or pair with each other, which facilitates the ligation, where the enzyme ligase forms a covalent bond between the vector and the insert, creating a new plasmid with the desired DNA sequence.

Most sequences are inserted into cloning vectors using the process of ligation. First, both the insert and the cloning vectors are digested with restriction enzymes. This digestion process produces either blunt ends or sticky ends, depending on the type of restriction enzyme used. After digestion, the insert and the vector are joined together using an enzyme called ligase, which creates a covalent bond between the two fragments. The resulting recombinant DNA molecule can then be used for further analysis or expression.