Culture Collections

COFUN - Aspergillus fumigatus gene-wide knock out collection

 

 COFUN

 

COFUN is a Wellcome Trust funded project undertaken by the University of Manchester that aims to generate knockout mutants for all of the coding genes in the human pathogen Aspergillus fumigatus. In total around 10,000 strains will be generated for which Culture Collections are the sole distributor.

The first sub-library consists of >400 transcription factor null mutants in the form of a panel (frozen). The transcription factor knockout strains incorporate a class III restriction site on one flank providing the capability to perform rapid quantitative parallel fitness analysis (the methodologies for this can be supplied on request).

 

NCPF No         Gene Knocked Out Format                 Price          
NCPF 7922 Transcription Factor DNA panel (frozen) (5x96 well) £1000

 

You can read more about the COFUN project in the COFUN leaflet

To keep up to date with the development of this resource and the release of libraries of kinase and phosphatase knockout mutants, follow us on Twitter @TheNCPF and sign-up to recieve updates from NCPF by email.

 

Handling Strains from the Transcription Factor Knock-Out Library

Material Safety Data Sheet For Aspergillus Fumigatus Transcription Factor KO Collection

Brief instructions on handling strains from the TF KO library.

The TFKO library is shipped in 50ul aliquots in 96 well plates. The strain identifiers are given in file COFUN004. All strains have been generated according to the overview document COFUN001. All knockout mutants have been purified by selection on hygromycin in SAB agar (Thermo Scientific CM0041 ) at 200 mg/L.

We would strongly suggest that before you use the isolates that you streak each strain to purity. Although we take all efforts here to avoid contamination working with large numbers of strains can be a challenge and we cannot absolutely guarantee that there has been no cross contamination.

 

Streaking strains to purity:

We would suggest that for each strain, a streak plate is prepared by plating 1µl of each spore stock on a selective plate (either SAB or any other suitable complete media; e.g. Aspergillus complete media) containing hygromycin at 200 mg/L. Incubate for 48 hours at 37˚C. Take conidia from a single colony and inoculate a small (12.5 cm2) tissue culture flask (must have a vented cap) with SAB or your complete media of choice containing hygromycin at 200 mg/L. Incubate for 3-5 days at 37˚C. Harvest spores into PBS containing 0.01% tween20. For long term storage, strains should be stored in solution containing glycerol (minimum 10%) at -80 ˚C.

 

Notes on strain validation:

Strains have been validated according to the process outlined in COFUN003. For the avoidance of doubt, single copy integration has not been verified by Southern blot. We strongly advise that the validity of any strains you work on is verified using your internal methods. The sequences of the primers used to generate the knockout cassettes are given in COFUN002 for your information.

 

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