DOE Human Genome Program Contractor-Grantee Workshop VIII
February 27-March 2, 2000  Santa Fe, NM

99. The RIKEN Mouse Full-Length cDNA Encyclopedia

Piero Carninci, Kazuhiro Shibata, Masayoshi Itoh, Hideaki Konno, Jun Kawai, Yuko Shibata, Yuichi Sugahara, T. Endo, Y. Ozawa, Yoshifumi Fukunishi, Atsushi Yoshiki, M. Kisakabe, Masami Muramatsu, Yasushi Okazaki, and Yoshihide Hayashizaki

Genome Science Laboratory, RIKEN, Tsukuba Life Science Center, 3-1-1 Koyadai, Japan

carninci@rtc.riken.go.jp

We report the ongoing efforts to prepare the mouse full-length cDNA Encyclopedia. We have, to the date of the submission of this abstract, constructed more than 150 full-length cDNA libraries. Libraries were prepared with the CAP trapper full-length cDNA selection method coupled to the trehalose-thermoactivated reverse transcriptase, in order to clone long full-length cDNAs. Additionally, in most libraries, cDNAs were normalized and subtracted to isolate rarely expressed full-length cDNAs. To date, we have clustered 100,907 3' end sequences containing at least half of all mouse full-length cDNAs in 690,130 successful sequencing reactions from 5'-end validated libraries. We have also established technology and mRNA resources to collect the majority of remaining mouse expressed sequences.

We will discuss about library preparation and tissue selection strategy, quality of cDNA libraries in terms of complexity and full-length cDNAs presence and coverage of mouse genes by our clones.