Patent ID: 6586180
Filing Date: 2003-07-01
Classification: C12N

Abstract:
A method for generating an oligonucleotide library targeted to a selected RNA transcript comprising:(a) preparing a double stranded cDNA, comprising a first end, a second end, and a central site thereof, from the selected RNA transcript and cloning the cDNA in a cloning vector comprising a promoter configured such that an antisense transcript of the cDNA is synthesized upon transcription mediated by the promoter, resulting in a cloned cDNA; (b) creating a plurality of deletion derivatives of said cloned cDNA by exonuclease resection thereof, wherein each of said plurality of deletion derivatives has a deletion extending from said first end towards the central site of the cloned cDNA such that the plurality of deletion derivatives comprises a deletion library comprising deletions that extend serially into the cDNA; and (c) reducing the size of the cDNA contained in the deletion library to a preselected size by removing a portion of the cDNA from the second end thereof to result in a fragment library; (d) inserting a catalytic core into the central site of the cDNA in the fragment library, thereby obtaining the oligonucleotide library.