What enables a PCR-amplified gene to be inserted into a cloning vector?
A) The PCR reaction adds sticky ends onto the amplified DNA, allowing ligation to a cloning vector.
B) Restriction enzyme sites are added to the 5' ends of both primers; therefore, digestion of the PCR-amplified gene allows ligation to a cloning vector cut with the same restriction enzyme.
C) The PCR-amplified gene is sequenced to locate restriction sites in order to determine which restriction enzymes to use for cloning.
D) The PCR-amplified gene is cut with one restriction enzyme, then ligated to a cloning vector cut by a different restriction enzyme.
E) The cloning vector is added to the PCR reaction, and once the gene is amplified, it immediately ligates to the cloning vector.

Question

Quizplus · Accepted Answer

Restriction enzyme sites are often added to the 5' ends of primers used in PCR. This allows the PCR product to be cut with the same restriction enzyme as the cloning vector, facilitating the ligation of the PCR-amplified gene into the vector.

What Enables a PCR-Amplified Gene to Be Inserted into a Cloning