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Nucleic Acids Research, Vol 24, Issue 17 3474-3475, Copyright © 1996 by Oxford University Press

ARTICLES

A positive screen for cloning PCR products

P Keese and L Graf
CSIRO, Division of Plant Industry, Canberra, Australia.

We have developed a positive screen for cloning PCR products based on translational activation of lacZ. A vector with a translationally deficient lacZ alpha gene has been made by deletion of the Shine- Dalgarno sequence and initiation codon. The Shine-Dalgarno sequence and initiation codon are incorporated into one of the PCR primers to allow complementation by the PCR product of the inactive lacZ alpha gene, which results in blue transformed bacterial colonies. This screen allows more efficient detection of clones containing inserts made by PCR.
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