| Literature DB >> 14501067 |
Bradley J Till1, Trenton Colbert, Rachel Tompa, Linda C Enns, Christine A Codomo, Jessica E Johnson, Steven H Reynolds, Jorja G Henikoff, Elizabeth A Greene, Michael N Steine, Luca Comai, Steven Henikoff.
Abstract
Targeting-induced local lesions in genomes (TILLING) is a general strategy for identifying induced point mutations that can be applied to almost any organism. Here, we describe the basic methodology for high-throughput TILLING. Gene segments are amplified using fluorescently tagged primers, and products are denatured and reannealed to form heteroduplexes between the mutated sequence and its wild-type counterpart. These heteroduplexes are substrates for cleavage by the endonuclease CEL I. Following cleavage, products are analyzed on denaturing polyacrylamide gels using the LI-COR DNA analyzer system. High-throughput TILLING has been adopted by the Arabidopsis TILLING Project (ATP) to provide allelic series of point mutations for the general Arabidopsis community.Entities:
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Year: 2003 PMID： 14501067 DOI： 10.1385/1-59259-413-1:205
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745