Xenopus
tropicalis EST Clones
Please note that the clone(s) being sent have tested
negative in our phage contamination assay. However these clones should
still be handled with care as no phage assay can be guaranteed to be 100%
accurate.
WHAT TO DO WITH YOUR CLONES WHEN YOU RECEIVE
THEM
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.
A selection of at least 10 single colonies should be picked and grown
in LB broth containing ampicillin + 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.
Clone Labeling and Nomenclature:
TEgg - clone from the X.tropicalis egg cDNA library.
The correct nomenclature of clone 35P4 from this library is TEgg035p04
TGas - clone from the X.tropicalis gastrula cDNA
library.
The correct nomenclature of clone 87J8 from this library is TGas087j08
TNeu - clone from the X.tropicalis neurula cDNA library.
The correct nomenclature of clone 131B12 from this library is TNeu131b12
TTpa - clone from the X.tropicalis tadpole cDNA library.
The correct nomenclature of clone 140f12 from this library is TTpa140f12
THda - clone from the X.tropicalis tailbud head cDNA library.
The correct nomenclature of clone 150b06 from this library is THda150b06
TTba - clone from the X.tropicalis tailbud cDNA library.
The correct nomenclature of clone 155c07 from this library is TTba155c07
Vector:
The vector used is pCS107 which was constructed in the Richard Harland's
laboratory
(http://mcb.berkeley.edu/labs/harland/).
For vector map see http://mcb.berkeley.edu/labs/harland/media/pdfs/CS107.pdf
For polylinker details see http://mcb.berkeley.edu/labs/harland/media/pdfs/CS105,107polylinker.pdf
Subsequent Publication:
In subsequent publications or database entries the clone must be referred
to by the full clone ID (e.g. TNeu128k06 is correct, 128k06 is incorrect).
In addition, please acknowledge the originators of the clone and reference Gilchrist et al., 2004*,
together with Geneservice for supplying the clone.
*Gilchrist, M., Zorn, A. M., Voigt, J, Smith, J.C., Papalopulu,
N. and Amaya, E. (2004). Defining a large set of full length
clones from a Xenopus tropicalis EST project. Dev.
Biol. 271: 498-516.
Version 2.0 25th May 2004
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