International Mammalian Genome Society


The 13th International Mouse Genome Conference
October 31-November 3, 1999

Table of Contents * Structure * Bioinformatics * Sequence * Mapping * New Tools * Gene Discovery * Developmental * Mutagenesis * Functional Genomics

I13 5'-end Analysis for Cap-Trapper Library

Yuichi Sugahara1,2, Carninci Piero1 and Yoshihide Hayashizaki1,2. 1Laboratory for Genome Exploration Research Project, Genomic Sciences Center (GSC) and Genome Science Laboratory, Tsukuba Life Science Center, the Institute of Physical and Chemical Research (RIKEN); 2CREST, Japan Science and Technology Corporation (JST)

To enhance the utility of laboratory mouse system and to facilitate the rapid assay of gene function, we are collecting an entire set of mouse full-length cDNA by single pass sequencing. To efficiently collect full-length cDNA clone, the high-quality cDNA library is the most critical. In recent years, we have been developing the full-length cDNA library construction method by using the biotinilation of cap structure, called cap-trapper, that is couples with a treatment to increase the reverse transcriptase efficiency at high temperature by the addition of trehalose. In this work, we evaluated the quality of cap-trapper library by comparing the 5 end of cDNA clones with those of public database. Our analysis showed that 63% of cDNA clones in the cap-trapper libraries had same or 5 extended sequence and 90% of clones maintained the coding sequence. These results indicate that the cap-trapper library is a promising tool to collect the full-length cDNA in large-scale EST projects.

 


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