GPCR clone set

Please note that the clone(s) being sent have tested negative in our T1 phage contamination assay. However these clones should still be handled with care as no phage assay can be guaranteed to be 100% reliable.

THE DISPATCH NOTE SHOWS THE ORIGINAL CLONE ID ORDERED AND ALSO THE ALIAS (PLATE AND WELL NUMBER ). THIS IS WHAT IS SHOWN ON THE CLONE LABEL. PLEASE KEEP THE DISPATCH NOTE FOR FUTURE REFERENCE. If you wish to translate from our plate to the original IMAGE ID, please use our plate conversion tool. Entering the ID as eg BF1

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/


Please go to the following website for full listing of available clones - http://www.imageconsortium.org/ftp/rearrayed_plates/IRBF-IRBI.preSV.data 

 

Acknowledgement

Please acknowledge Guthrie cDNA Resource Center (http://www.guthrie.org/) in any resulting publications.