Worm Breeder's Gazette 17(1): 30 (October 1, 2001)

These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.

Gene Knockout Methods Available Online

Sheldon McKay1, Mark Edgley1, Erin Gilchrist1, Steven Jones2, Don Moerman1,3

1 Caenorhabditis elegans Reverse Genetics Core Facility, Biotechnology Laboratory, University of British Columbia, Vancouver, BC
2 Genome Sequence Centre, British Columbia Cancer Agency, Vancouver, BC
3 Department of Zoology, University of British Columbia, Vancouver, BC

The Vancouver node of the C. elegans gene knockout consortium has made information on the gene knockout process available on the web (http://ko.cigenomics.bc.ca/protocols.html) for researchers who wish to perform gene knockout experiments in their own labs.  The time-line and methods for the gene knockout process is described, from mutagenesis to isolation of individual mutations.  The information also includes a detailed description and experimental protocols for PCR screening of mutagenized libraries using the "poison primer" technique.  Our library construction methods are modeled after the Moulder and Barstead protocol (http://www.mutantfactory.ouhsc.edu/protocols.asp), while our mutagenesis protocol is modified from the method developed by Shohei Mitani.

We have also released an updated version of a web-accessible program that uses information in ACeDB to design PCR primers for gene knockout experiments (http://ko.cigenomics.bc.ca/oligos.shtml) and a searchable database of pre-designed knockout primer sets for standard and "poison primer" PCR screening (http://ko.cigenomics.bc.ca/oligodb.shtml).