Question

Choose the correct statement with respect to the self priming method of cDNA synthesis.

(a) It is less preferred than RNaseH method

(b) A hairpin structure is formed with guarantee

(c) The sequence corresponding to the 5’ end is lost

(d) Reverse transcriptase is not used

I got this question in an interview.

The query is from cDNA Libraries, Polyadenylated RNA and cDNA Synthesis topic in section Libraries Construction of Genetic Engineering

Answer 1

Correct answer is (c) The sequence corresponding to the 5’ end is lost

The explanation: The self priming method is based on the formation of a hairpin structure. But there are only chances and no guarantee of formation of hairpin structure. In the first strand synthesis, reverse transcriptase is used. The sequence corresponding to the 5’ end is lost and in cDNA large deletions are observed. This method is less preferred than RNaseH method.