What is the most logical sequence of steps for splicing foreign DNA into a plasmid and inserting the plasmid into a bacterium?
I. Transform bacteria with a recombinant DNA molecule.
II. Cut the plasmid DNA using restriction enzymes (endonucleases).
III. Extract plasmid DNA from bacterial cells.
IV. Hydrogen-bond the plasmid DNA to non-plasmid DNA fragments. V. Use ligase to seal plasmid DNA to non-plasmid DNA.
V. Use ligase to seal plasmid DNA to non-plasmid DNA.

• A. II, III, V, IV, I
• B. III, II, IV, V, I
• C. III, IV, V, I, II
• D. IV, V, I, II, III

Answer: (B)

Verified step by step guidance

1. Step 1: Begin by extracting plasmid DNA from bacterial cells (Step III). This involves isolating the plasmid DNA, which will serve as the vector for inserting foreign DNA.
2. Step 2: Use restriction enzymes (endonucleases) to cut the plasmid DNA at specific recognition sites (Step II). This creates openings in the plasmid where foreign DNA can be inserted.
3. Step 3: Allow the plasmid DNA to hydrogen-bond with non-plasmid DNA fragments (Step IV). This step involves complementary base pairing between the sticky ends of the plasmid and the foreign DNA.
4. Step 4: Use DNA ligase to seal the plasmid DNA to the non-plasmid DNA fragments (Step V). Ligase catalyzes the formation of phosphodiester bonds, creating a stable recombinant DNA molecule.
5. Step 5: Transform bacteria with the recombinant DNA molecule (Step I). This involves introducing the recombinant plasmid into bacterial cells, where it can replicate and express the foreign DNA.