Inverse Polymerase Chain Reaction
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Last Update: 2020-11-28
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Source: Internet
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Author: User
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Since the first report on c
DNA
cloning in 1972 (
1
), this technology has been developed into a powerful and universal tool in the isolation, characterization, and analysis of both eukaryotic and prokaryotic genes. But the conventional methods of
cDNA
cloning require much effort to generate a library that is packaged in phage or plasmid and then survey a large number of recombinant phages or plasmids. There are three major limitations in those methods. First, substantial amount (at least 1 �g) of purified mRNA is needed as starting material to generate libraries of sufficient diversity (
2
). Second, the intrinsic difficulty of multiple sequential enzymatic reactions required for cDNA cloning often leads to low yields and truncated clones (
3
). Finally, screening of a library with hybridization technique is time consuming.
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