A positive screen for cloning PCR products | Zendy

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A positive screen for cloning PCR products

Author(s) -

Paul Keese

Publication year - 1996

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/24.17.3474

Subject(s) - biology , start codon , cloning (programming) , complementation , gene , microbiology and biotechnology , lac operon , genetics , stop codon , polymerase chain reaction , plasmid , messenger rna , computer science , phenotype , programming language

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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