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The Illumina-Solexa Sequencing Protocol for Bacterial Genomes

  • Zhenfei Hu
  • Lei Cheng
  • Hai WangEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1231)

Abstract

Based on reversible dye-terminators technology, the Illumina-solexa sequencing platform enables rapid sequencing-by-synthesis (SBS) of large DNA stretches spanning entire genomes, with the latest instruments capable of producing hundreds of gigabases of data in a single sequencing run. Illumina’s NGS instruments powerfully combine the flexibility of single reads with short- and long-insert paired-end reads, and enable a wide range of DNA sequencing applications. Here, we describe the paired-end library preparation with an average insert size of 470 bp, 2 kbp, and 6 kbp, together with the DNA cluster generation and sequencing procedure of E. coli O104:H4 genome on Illumina Hiseq 2000 platform.

Key words

Illumina Solexa Hiseq Reversible dye-terminators Sequencing by synthesis Single reads Paired-end reads Library preparation Cluster generation 

Notes

Acknowledgements

This work was supported by E. coli O104:H4 Genome Analysis Crowd-Sourcing Consortium.

References

  1. 1.
    Rohde H, Qin J, Cui Y (2011) Open-source genomic analysis of Shiga-toxin-producing E. coli O104:H4. N Engl J Med 365:718–724CrossRefPubMedGoogle Scholar
  2. 2.
    Frank C, Werber D, Cramer JP et al (2011) Epidemic profile of Shiga-toxin-producing Escherichia coli O104:H4 outbreak in Germany—preliminary report. N Engl J Med 365:1771–1780CrossRefPubMedGoogle Scholar
  3. 3.
    Robert Koch Institute (2011) Information up-date on EHEC/HUS outbreak. http://www.rki.de/nn_217400/EN/Home/PM082011.html

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.BGIShenzenChina

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