Which of the Following is NOT a Step in the Southern

Question

Which of the following is NOT a step in the Southern blotting procedure?
A)Digestion of the DNA with a restriction enzyme
B)Ligation of the DNA into a vector
C)Separation of the DNA fragments on a gel
D)Transfer of the DNA fragments to a nitrocellulose membrane
E)Hybridization of the membrane-bound DNA with a labeled probe

Accepted Answer

Ligation of DNA into a vector is not a step in Southern blotting. The steps in Southern blotting are digestion of the DNA with a restriction enzyme, separation of the DNA fragments on a gel, transfer of the DNA fragments to a nitrocellulose membrane, and hybridization of the membrane-bound DNA with a labeled probe. The purpose of Southern blotting is to identify specific DNA sequences within a sample.