454 sequencing put to the test using the complex genome of barley
2006

Testing 454 Sequencing on Barley Genome

Sample size: 4 publication Evidence: moderate

Author Information

Author(s): Wicker Thomas, Schlagenhauf Edith, Graner Andreas, Close Timothy J, Keller Beat, Stein Nils

Primary Institution: Institute of Plant Biology, University of Zürich

Hypothesis

Can 454 sequencing be an efficient and cost-effective alternative to traditional ABI-Sanger sequencing in repetitive genomes?

Conclusion

454 pyrosequencing allows rapid and cost-effective sequencing of gene-containing portions of large and complex genomes.

Supporting Evidence

  • 454 sequencing provided a much more even coverage of the BAC clones than ABI-Sanger sequencing.
  • All gene-containing regions were covered efficiently and at high quality with 454 sequencing.
  • The combination of 454 sequencing with ABI-Sanger sequencing can result in high-quality finished genomic sequences.

Takeaway

This study shows that a new sequencing method can quickly and cheaply read the important parts of big plant genomes, like barley.

Methodology

The study used 454 sequencing technology to sequence four barley BAC clones and compared the results to ABI-Sanger sequencing.

Limitations

The assembly of repetitive sequences was problematic, and finished BAC sequences were not achieved solely with 454 technology.

Digital Object Identifier (DOI)

10.1186/1471-2164-7-275

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