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Tilling by Sequencing

  • Helen Tsai
  • Kathie Ngo
  • Meric Lieberman
  • Victor Missirian
  • Luca ComaiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1284)

Abstract

TILLING is a method to find mutations in a gene of interest by scanning amplicons from a mutagenized population for sequence changes, commonly a single nucleotide. In the past 5 years, mutation detection by sequencing has become increasingly popular. This chapter details the experimental flow for TILLING-by-Sequencing, highlighting the critical steps involved in tridimensional pooling of genomic DNA templates, preparation of libraries for high-throughput sequencing, and bioinformatic processing of the sequence data.

Key words

Mutation discovery Pooling High-throughput sequencing Bioinformatics 

Notes

Acknowledgments

This work was supported by the National Science Foundation (Plant Genome award no. DBI–0822383).

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Helen Tsai
    • 1
  • Kathie Ngo
    • 1
  • Meric Lieberman
    • 1
  • Victor Missirian
    • 1
  • Luca Comai
    • 1
    Email author
  1. 1.Department of Plant Biology and Genome CenterUniversity of California at DavisDavisUSA

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