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Related Experiment Videos

High-throughput TILLING for functional genomics.

Bradley J Till1, Trenton Colbert, Rachel Tompa

  • 1BasicSciences Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 23, 2003
PubMed
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Targeting-Induced Local Lesions in Genomes (TILLING) is a high-throughput method to find induced point mutations. This technique uses endonuclease cleavage and gel analysis to identify genetic variations in organisms like Arabidopsis.

Area of Science:

  • Genetics
  • Molecular Biology
  • Genomics

Background:

  • Targeting-Induced Local Lesions in Genomes (TILLING) is a versatile strategy for discovering induced point mutations.
  • This method is applicable to a wide range of organisms for genetic research.

Purpose of the Study:

  • To describe the fundamental methodology for performing high-throughput TILLING.
  • To detail the process of identifying point mutations using TILLING.

Main Methods:

  • Gene segments are amplified with fluorescently tagged primers.
  • Denatured and reannealed products form heteroduplexes for CEL I endonuclease cleavage.
  • Cleaved products are analyzed on denaturing polyacrylamide gels using a LI-COR DNA analyzer.

Main Results:

Related Experiment Videos

  • The described methodology enables efficient high-throughput screening for induced point mutations.
  • The Arabidopsis TILLING Project (ATP) utilizes this method to generate allelic series of mutations.

Conclusions:

  • High-throughput TILLING provides a powerful tool for genetic variation discovery.
  • This approach facilitates the study of gene function through the provision of diverse mutant alleles to the research community.