Team:Grenoble/Biology/Protocols/Double

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iGEM Grenoble 2012

Project

Double mutant construction

Goal

Obtain double gene knockout mutant strains of E. coli from single mutant strains (Keio collection).

Protocol

From the single mutant strains, the first step was to remove the Kan cassette in order to create the receiver strain. The second step is using the P1 bacteriophage to transfer a Kan cassette from another bacteria to this receiver stain, in order to replace the second gene.