GENE LIBRARY

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GENE LIBRARY

To clone a DNA sequence that codes for the required gene product, The gene has to be removed fro the organism & cloned in a vector molecule. So libraries of such cloned fragments are made.
A gene library is a random collection of cloned fragments in a suitable vector that ideally includes all the genetic information of that species. It is also called as shot gun library.

The identification & isolation of the desired gene/ DNA fragment/DNA insert to be cloned is an important step in gene cloning. The desired gene can be obtained from the following:

COMPLEMENTARY (cDNA) LIBRARIES

GENOMIC DNA LIBRARY

Chemical/enzymatic synthesis

Amplification through PCR

CHEMICAL/ENZYMATIC SYNTHESIS OF DNA

This method uses the bacterial enzymes polynucleotide phosphorylase which is specific for ribo nucleotides, but also polymerizes deoxyribonucleotides at a sloe rate when Mn ²⁺ replaces Mg2+ in the reaction mixture. It uses Deoxyribonucleoside diphosphates as substrate & does not require a template but a primer to which enzyme adds 1 or 2 nucleotides at a time. The reactions are limited & hard to control.