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WHAT TO DO WITH YOUR CLONES WHEN YOU RECEIVE THEM
Plates- contain LB broth with 8% glycerol and ampicillin (50 µg/ml)
are sent on Dry Ice. These should be stored at -70°C.
Individual clones have been streaked onto LB agar containing ampicillin
(50 µg/ml). 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.
A selection of at least 10 single colonies should be picked and grown
in LB broth containing 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.
The GPCR clone collection has been produced by Guthrie cDNA Resource
Center (http://www.guthrie.org/)
which is a non-profit organization. The cDNA clones have been produced
in pcDNA3.1+ and pDNR-1r vectors. Please click on the links below for
vector information:
pcDNA3.1 http://www.invitrogen.com/content/sfs/vectors/pcdna3.1+.pdf
pDNR-1r http://www.clontech.com/techinfo/vectors/vectorsD/pDNR-1r.shtml
Please go to the following WWW site for full listing of available clones
- http://image.llnl.gov/image/html/iresources.shtml
Acknowledgement
Please acknowledge Guthrie cDNA Resource Center (http://www.guthrie.org/)
in any resulting publications.
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