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Comparative Lesions Analysis Through a Targeted Sequencing Approach
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Tilling by sequencing.

Helen Tsai1, Kathie Ngo, Meric Lieberman

  • 1Department of Plant Biology and Genome Center, University of California at Davis, One Shields Avenue, Davis, CA, 95616, USA.

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|March 12, 2015
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Summary
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Targeting Induced Local Lesions IN Genomes (TILLING) by Sequencing efficiently detects single nucleotide mutations. This method streamlines mutation discovery by combining DNA pooling, library preparation, and bioinformatics for high-throughput sequencing analysis.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Bioinformatics

Background:

  • TILLING (Targeting Induced Local Lesions IN Genomes) is a valuable method for identifying mutations in specific genes.
  • Mutation detection using DNA sequencing has gained significant traction in recent years.
  • Efficiently screening mutagenized populations for genetic variations is crucial for research.

Purpose of the Study:

  • To provide a detailed experimental workflow for TILLING-by-Sequencing.
  • To highlight key steps in the process, from DNA pooling to data analysis.
  • To facilitate the adoption and understanding of this high-throughput mutation detection technique.

Main Methods:

  • Utilizing tridimensional pooling of genomic DNA templates to organize samples efficiently.
  • Preparing DNA libraries specifically designed for high-throughput sequencing platforms.
  • Implementing bioinformatic pipelines for processing and analyzing large-scale sequence data.

Main Results:

  • The chapter outlines a comprehensive protocol for TILLING-by-Sequencing.
  • Critical steps for successful implementation are clearly defined.
  • The workflow integrates sample preparation, sequencing, and computational analysis.

Conclusions:

  • TILLING-by-Sequencing offers a powerful approach for rapid and accurate mutation discovery.
  • The described methodology enables efficient screening of mutagenized populations.
  • This technique is essential for genetic research and functional genomics studies.