TRANSCRIPTOME 2002: From Functional Genomics to Systems
Generating and Sequencing Full-length cDNAs of Novel Human Genes Within the German cDNA Consortium
Ruth E. Wellenreuther, Stefan Wiemann, Daniela Heiss, Nina Claudino, Annemarie Poustka, Department of Molecular Genome Analysis, German Cancer Research Center, Im Neuenheimer Heidelberg, GERMANY
We generate human cDNA libraries that are enriched in full length clones i.e. from the translation start to the poly A tail. These libraries are used for a) systematic sequencing within the cDNA consortium of the Genome Project aiming at the identification and analysis of as many new genes as possible and b) for screening to isolate full length clones of partial genes. Libraries are created by directional cloning of cDNA's into plasmid vectors. Full-length enrichment is achieved via Clontech's SMART technology. This method is PCR-based, and in our modified strategy, we amplify and clone selective size windows of the cDNA fraction above 3 kb.Clones from the libraries generated within this project are the major source for the cDNA sequencing effort of the German Human Genome Project that is carried out by a consortium of nine laboratories. Over 900 full-length cDNAs of novel human genes could be identified by the consortium within the last 4 years. The clones generated in this project are available through the Resource Center of the German Human Genome Project (www.rzpd.de).
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