Applications of Ultra-high-Throughput Sequencing

  • Samuel Fox
  • Sergei Filichkin
  • Todd C. Mockler
Part of the Methods in Molecular Biology™ book series (MIMB, volume 553)

Abstract

The genomics era has enabled scientists to more readily pose truly global questions regarding mutation, evolution, gene and genome structure, function, and regulation. Just as Sanger sequencing ushered in a paradigm shift that enabled the molecular basis of biological questions to be directly addressed, to an even greater degree, ultra-high-throughput DNA sequencing is poised to dramatically change the nature of biological research. New sequencing technologies have opened the door for novel questions to be addressed at the level of the entire genome in the areas of comparative genomics, systems biology, metagenomics, and genome biology. These new sequencing technologies provide a tremendous amount of DNA sequence data to be collected at an astounding pace, with reduced costs, effort, and time as compared to Sanger sequencing. Applications of ultra-high-throughput sequencing (UHTS) are essentially limited only by the imaginations of researchers, and include genome sequencing/resequencing, small RNA discovery, deep SNP discovery, chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) coupled with sequence identification, transcriptome analysis including empirical annotation, discovery and characterization of alternative splicing, and gene expression profiling. This technology will have a profound impact on plant breeding, biotechnology, and our fundamental understanding of plant evolution, development, and environmental responses. In this chapter, we provide an overview of UHTS approaches and their applications. We also describe a protocol we have developed for deep sequencing of plant transcriptomes using the Illumina/Solexa sequencing platform.

Key words

Ultra-high-throughput DNA sequencing HTS UHTS microread sequencing transcriptome 454 Illumina Solexa SOLiD 

Notes

Acknowledgements

We thank Rongkun Shen, Henry Priest, Scott Givan, Chris Sullivan, and Doug Bryant for their contributions to our studies involving Illumina-based transcriptome sequencing. We would also like to thank Alice Barkan, Todd Castoe, Todd Michael, and Laurence von Kalm for their helpful comments on this chapter. This work was supported by Oregon State University and by grant ARF4435 from the Oregon Agricultural Research Foundation.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Samuel Fox
    • 1
  • Sergei Filichkin
    • 1
  • Todd C. Mockler
    • 1
  1. 1.Department of Botany and Plant Pathology and Center for Genome Research and BiocomputingOregon State UniversityCorvallisUSA

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