Recombineering Reagent

Protocols

Below you'll see a short description of each protocol and a link to a PDF file for download.

Recombineering protocol 1
General protocol for how to use the DY380 strain for recombineering. Included is how to perform the heat-shock, how to make electrocompetent cells, and how to prepare the DNA for recombineering experiments.

Recombineering protocol 2
Explains how to make a conditional knock-out vector using the EL350 strain. Basically, as a piece of genomic DNA is retrieved from a BAC into a plasmid vector followed by two rounds of targeting using loxP-flanked (floxed) neo cassettes.

Recombineering protocol 3
Explains how to use the galK positive and counterselection scheme to make essentially any BAC modification (e.g. point mutations, deletions, and insertions). [Updated on June 24, 2005]

Recombineering protocol 4
Explains how to design a good strategy for a conditional knock-out project, including several important issues you should consider in the design process.