Reverse transcriptase and terminal deoxynucleotidyl transferase are used in gene cloning to
A) remove single stranded DNA and repair the restriction enzyme site after transformation respectively.
B) make cDNA and create sticky ends on the cDNA and the staggered ends of the cleaved restriction site.
C) repair the restriction enzyme site before transformation.
D) repair the restriction enzyme site after transformation.
E) remove the sticky ends prior to the DNA ligase step.

Question

Quizplus · Accepted Answer

Reverse transcriptase is used to make complementary DNA (cDNA) from an RNA template, which can then be cloned into a vector using restriction enzymes. Terminal deoxynucleotidyl transferase (TdT) is used to add non-template nucleotides to the 3' ends of the cDNA, creating "sticky ends" that can hybridize with the staggered ends of the cleaved restriction site. This results in a more stable ligation and increases the efficiency of cloning.

Reverse Transcriptase and Terminal Deoxynucleotidyl Transferase Are Used in Gene