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ZGC (Zebrafish Gene Collection)

ZGC (Zebrafish Gene Collection)

The Zebrafish Gene Collection (ZGC) project is an NIH initiative which was launched in April 2002, as a subproject of the existing Mammalian Gene Collection. The general approach is to acquire or construct high-quality cDNA libraries. The libraries are screened by sequencing the 5' end of the clones. Putative full open reading frames are identified, and finally, selected clones are full-length sequence verified. The goal of the project is to make available a complete set of these full-length cDNA clones in Zebrafish.

IRBO ZGC Zebrafish full length amp cDNA
IRCX Zebrafish predicted full length kan cDNA
IRCY Zebrafish PCR rescued genes kan cDNA
IRBV ZGC Zebrafish verified full-length cam cDNA

Protocols / Clone Handling

IRBO (Zebrafish)
The clones have been streaked onto LB agar containing 50µg/ml ampicillin. Please store them at 4°C (not in a freezer).
As soon as possible, re-streak the clones onto the same medium and incubate overnight at 37°C to obtain single colonies.

IRBV (Zebrafish)
The clones have been streaked onto LB agar containing 57µg/ml chloramphenicol. Please store them at 4°C (not in a freezer).
As soon as possible, re-streak the clones onto the same medium and incubate overnight at 30°C to obtain single colonies.

IRCX and IRCY (Zebrafish), kanamycin, 30 μg/ml
A selection of at least 10 single colonies should be picked and grown in LB broth containing the appropriate antibiotic + 8% glycerol for subsequent freezing at -70°C. These frozen stocks can then be re-grown for plasmid isolation and purification.
Streaking out for single colonies is essential in case of contamination or cross-contamination, or if modifications of the original clone have taken place as the library has been replicated. If there is a problem, the more single colonies you streak out, the more likely it is that you can retrieve the original construct. You can also isolate the original clone from a contaminated sample by following this procedure.

References

MGC (Mammalian Gene Collection) Program Team (2002). Generation and Initial Analysis of more than 15000 Full-Length Human and Mouse cDNA sequences. PNAS 99(26): 16899-16903


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