NOVEL HIGH-THROUGHPUT SYSTEM FOR FUNCTIONAL GENOMICS USING UNIDIRECTIONAL ANTISENSE CDNA LIBRARY

摘要

PURPOSE: Provided is a novel high-throughput system for functional genomics using unidirectional and subtracted antisense cDNA library, thereby analyzing novel genomics. CONSTITUTION: The method for constructing unidirectional antisense cDNA library comprises the steps of: separating RNA from a specific or disease related cell line or tissue; performing reverse transcription using the RNA as a template to synthesize cDNA strands therefrom; linking a restriction enzyme adaptor to ends of the synthesized cDNA; cloning cDNA fragment into a phagemid vector treated with the same restriction enzyme above to obtain a recombinant phagemid; manufacturing a transformant using the recombinant phagemid; and producing single stranded antisense DNA by coinfecting the transformant and a helper phage to construct unidirectional antisense cDNA library.