Xenopus tropicalis cDNA Collections
The Xenopus tropicalis EST Project is a joint collaboration between the Sanger Institute and the Wellcome Trust/Cancer Research UK Gurdon Institute 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.crick.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. To search by gene name or blast click here.
If you require further information on the additional plates, or would like to become part of a "user" group, please contact https://www.crick.ac.uk/mike-gilchrist
For specific information concerning plate or clone ordering please contact us.
The Sanger Institute has Generated approximately 55,000 5' EST sequences from each X. tropicalis cDNA library.
The complete set of 9180 full length clones is contained in 24 384-well plates (Arraying details). Publications stemming from clones obtained from these libraries should reference Gilchrist et al.,2004.
- Identify unknown genes and map their position in a genome
- Genome map construction
- Characterization of expressed genes
Due to MTA requirements, individual clones and libraries should be ordered via our website.
Find the clone you are looking for by gene name or blast click here, or browse NCBI dbEST , and searched and ordered on our website below, using the clone id (e.g. CR589752.1).
The entire set can be ordered below:
||Xenopus tropicalis Full Length cDNA
The use of the resource is limited to research purposes. The collection is subject to the following MTA and our clone terms and conditions.
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.
Protocols / Clone Handling
The clones have been streaked onto LB agar containing 50g/ml ampicillin. Please store them at 4oC (not in a freezer).
As soon as possible, re-streak the clones onto the same medium and incubate overnight at 37oC 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 –70oC. 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.
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.
For further information and prices please contact us or call +44 (0)115 973 9012