Question

If hybridization is carried out with a clone having a sequence similar to that of the vector from which library has been constructed, then it is of no use. Choose the correct statement in respect to it.

(a) PCR amplification of the probe can be carried out

(b) Excision of the probe should be carried out which should be followed polyacrylamide gel electrophoresis

(c) If the vector sequence and the DNA probe are having similarities, some of the vector would be hybridized

(d) Probe should be in a vector that is having some sequence similarity with the library to be screened

The question was asked in homework.

My doubt is from Database Screening and Screening by Nucleic Acid Hybridisation topic in section Screening Libraries of Genetic Engineering

Answer 1

Right choice is (a) PCR amplification of the probe can be carried out

Easiest explanation: If hybridization is carried out with a clone having a sequence similar to that of the vector from which the library has been constructed then all the vector would hybridize. To make the sequence different, PCR amplification of the probe can be carried out. Also, excision of the probe can be carried out by using suitable restriction enzymes and it is followed by agarose gel electrophoresis. Or probe in a vector should not have any sequence similarity with the library to be screened.