Question

Arrange the following steps of site-directed mutagenesis in correct order :
A. Transformation of ds DNA into cells
B. Denaturation and annealing of oligonucleotide primers with mutation
C. Repair of nicks in mutated plasmid inside cells
D. Extension and incorporation of primers by using DNA polymerase
E. Digestion of unmutated parental DNA template with methylation specific nuclease and annealing of newly synthesized strands
Choose the correct answer from the options given below :

• A. \(B, D, E, A, C\)
• B. \(E, D, B, C, A\)
• C. \(C, B, E, D\)
• D. \(A, C, D, E, B\)

Hint:

Remember the key steps in site-directed mutagenesis: \[ \text{Primer Annealing} \rightarrow \text{DNA Synthesis} \rightarrow \text{Template Digestion} \rightarrow \text{Transformation} \]
• DpnI digests methylated parental DNA.
• Mutagenic primers introduce the desired mutation.

Answer 1

The Correct Option is A