High-throughput TILLING for functional genomics.

Bradley J Till; 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

doi:10.1385/1-59259-413-1:205

PubMed · 14501067

High-throughput TILLING for functional genomics.

Bradley J Till·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

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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.

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DNA, Plant Genes, Plant Genetic Techniques Mutagenesis Mutagens Nucleic Acid Heteroduplexes Plants Polymerase Chain Reaction Journal Article

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BibTeX RIS

Bradley J Till, 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. 2003. High-throughput TILLING for functional genomics.. https://doi.org/10.1385/1-59259-413-1%3A205

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