Non-mammalian clones

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

For specific information 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.