Skip to main content

OligoTag: A Program for Designing Sets of Tags for Next-Generation Sequencing of Multiplexed Samples

  • Protocol
  • First Online:
Data Production and Analysis in Population Genomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 888))

Abstract

Next-generation sequencing systems allow high-throughput production of DNA sequence data. But this technology is more adapted for analyzing a small number of samples needing a huge amount of sequences rather than a large number of samples needing a small number of sequences. One solution to this problem is sample multiplexing. To achieve this, one can add a small tag at the extremities of the sequenced DNA molecules. These tags will be identified using bioinformatics tools after the sequencing step to sort sequences among samples. The rules to apply for selecting a good set of tags adapted to each situation are described in this chapter. Depending on the number of samples to tag and on the required quality of assignation, different solutions are possible. The software oligoTag, a part of OBITools that computes these sets of tags, is presented with some example sets of tags.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. O’Brien HE, Parrent JL, Jackson JA et al (2005) Fungal community analysis by large-scale sequencing of environmental samples. Appl Environ Microbiol 7:5544–5550

    Article  Google Scholar 

  2. Briée C, Moreira D, López-García P (2007) Archaeal and bacterial community composition of sediment and plankton from a suboxic freshwater pond. Res Microbiol 158:213–227

    Article  PubMed  Google Scholar 

  3. Fierer N, Morse JL, Berthrong ST et al (2007) Environmental controls on the landscape-scale biogeography of stream bacterial communities. Ecology 88:2162–2173

    Article  PubMed  Google Scholar 

  4. Nicol GW, Leininger S, Schleper C, Prosser JI (2008) The influence of soil ph on the diversity, abundance and transcriptional activity of ammonia oxidizing archaea and bacteria. Environ Microbiol 10:2966–2978

    Article  PubMed  CAS  Google Scholar 

  5. Zinger L, Coissac E, Choler P, Geremia RA (2009) Assessment of microbial communities by graph partitioning in a study of soil fungi in two alpine meadows. Appl Environ Microbiol 75:5863–5870

    Article  PubMed  CAS  Google Scholar 

  6. Teixeira LCRS, Peixoto RS, Cury JC et al (2010) Bacterial diversity in rhizosphere soil from antarctic vascular plants of admiralty bay, maritime antarctica. ISME J 4:989–1001

    Article  PubMed  Google Scholar 

  7. Cronn R, Liston A, Parks M et al (2008) Multiplex sequencing of plant chloroplast genomes using solexa sequencing-by-synthesis technology. Nucleic Acids Res 36:e122

    Article  PubMed  Google Scholar 

  8. Hamady M, Walker JJ, Harris JK et al (2008) Error-correcting barcoded primers for pyrosequencing hundreds of samples in multiplex. Nat Methods 5:235–237

    Article  PubMed  CAS  Google Scholar 

  9. Cock PJA, Fields CJ, Goto N et al (2009) The sanger fastq file format for sequences with quality scores, and the solexa/illumina fastq variants. Nucleic Acids Res 38:1767–1771

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Eric Coissac .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media New York

About this protocol

Cite this protocol

Coissac, E. (2012). OligoTag: A Program for Designing Sets of Tags for Next-Generation Sequencing of Multiplexed Samples. In: Pompanon, F., Bonin, A. (eds) Data Production and Analysis in Population Genomics. Methods in Molecular Biology, vol 888. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-870-2_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-870-2_2

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-869-6

  • Online ISBN: 978-1-61779-870-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics