Xenopus tropicalis EST Clones
The Xenopus tropicalis EST Project is a
joint collaboration between the Sanger Institute and the Wellcome Trust/Cancer
Research UK Gurdon Institute (http://www.gurdon.cam.ac.uk//) with the aim of
supporting the X. tropicalis genetics and genomics effort. Source
BioScience LifeSciences are supplying clones from four Xenopus
(Silurana) tropicalis cDNA libraries:
X. tropicalis neurula cDNA library - TNeu
X. tropicalis gastrula cDNA library - TGas
X. tropicalis egg cDNA library - TEgg
X. tropicalis tadpole cDNA library - TTpa
X. tropicalis tailbud Head cDNA library - THda
X. tropicalis tailbud cDNA library - TTba
X. tropicalis Full length cDNA set - TFL
Clones are supplied individually or as a complete set of
full-length clones (Gilchrist et al., 2004*) (http://genomics.nimr.mrc.ac.uk/online/xt-fl-db.html).
Source BioScience LifeSciences are pleased to announce that
Mike Gilchrist of the Gurdon Institute, University of Cambridge,
has released four new full-length Xenopus tropicalis plates for us
to distribute. The 384-well plates are part of the Xt3 project and
have been designated plates 21-24, within the overall
collection.
If you require further information on the additional plates, or
would like to become part of a "user" group, please contact: m.gilchrist@nimr.mrc.ac.uk
Specifics concerning plate or clone ordering please contact
us.
The complete set of 9180 clones is contained in 24 384-well
plates ( Arraying details). Publications stemming from
clones obtained from these libraries should reference Gilchrist et
al., 2004.
Construction of the libraries is described in Gilchrist et al.,
2004. They were arrayed and sequenced by the Xenopus
tropicalis team (http://www.sanger.ac.uk/Projects/X_tropicalis/team.shtml).
Briefly, 5 ug of poly A+ RNA from each of the three embryonic
stages was oligo dT primed (Not1/Sal1 dT primer). After second
strand synthesis, EcoRI-SmaI adapters were ligated and the cDNA was
directionally cloned with EcoR1 at the 5' end and Not1 at the 3'
end. Each library started with 1 - 4 million original recombinants,
with an average insert size of about 1.5 kb and a range of 0.5 - 4
kb on 20 random clones. The percentage of empty vector is around
1%. The vector used is pCS107 (map, polylinker) and the host bacteria is XL1 blue for the gastrula and egg clones
and DH10B (Invitrogen) for the neurula and tadpole clones.
The Sanger Institute has generated approximately 55,000 5' EST
sequences from each X. tropicalis cDNA library.
*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.