Skip to main content

\(^\prime\)- and 3\(^\prime\)-RACE from LongSAGE Tags

  • Protocol
Serial Analysis of Gene Expression (SAGE)

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 387))

  • 648 Accesses

summary

Serial analysis of gene expression (SAGE) studies often yield numerous tags that cannot be mapped to known gene sequences. Intriguingly, these may represent unknown genes, unknown parts of genes, or transcript variants. In order to elucidate the origin of these tags, 3\(^\prime\)- and 5\(^\prime\)-rapid amplification of complementary DNA ends (RACE) reactions can be performed using primers identical or complementary to SAGE tags. This way, transcript fragments, or indeed the entire uncharacterized transcript, can be cloned and sequenced.

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 109.99
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

Reference

  1. Velculescu, V. E., Zhang, L., Vogelstein, B., and Kinzler, K. W. (1995) Serial analysis of gene expression. Science 270, 484–487.

    Article  CAS  PubMed  Google Scholar 

  2. Saha, S., Sparks, A. B., Rago, C., et al. (2002) Using the transcriptome to annotate the genome. Nat. Biotechnol. 20, 508–512.

    Article  CAS  PubMed  Google Scholar 

  3. Nielsen, K. L., Grønkjær, K., Welinder,K. G., and Emmersen,J. (2005) Global transcript profiling of potato tuber using LongSAGE. Plant Biotechnol. J. 3, 175–185.

    Article  CAS  PubMed  Google Scholar 

  4. Boheler, K. R. and Stern, M. D. (2003) The new role of SAGE in gene discovery. Trends Biotechnol. 21, 55–57.

    Article  CAS  PubMed  Google Scholar 

  5. Chen, J. J., Rowley, J. D., and Wang, S. M. (2000) Generation of longer cDNA fragments from serial analysis of gene expression tags for gene identification. Proc. Natl. Acad. Sci. USA 97, 349–353.

    Article  CAS  PubMed  Google Scholar 

  6. Chen, J., Lee, S., Zhou, G., and Wang, S. M. (2002) High-throughput GLGI procedure for converting a large number of serial analysis of gene expression tag sequences into 3\(^\prime\) complementary DNAs. Genes Chromosomes Cancer 33, 252–261.

    Article  CAS  PubMed  Google Scholar 

  7. Frohman, M. A., Dush, M. K., and Martin, G. R. (1988) Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc. Natl. Acad. Sci. USA 85, 8998–9002.

    Article  CAS  PubMed  Google Scholar 

  8. van den Berg, A., van der Leij, J. and Poppema, S. (1999) Serial analysis of gene expression: rapid RT-PCR analysis of unknown SAGE tags. Nucleic Acids Res. 27, e17.

    Article  PubMed  Google Scholar 

  9. Elhafi, G., Naylor, C. J., Savage, C. E., and Jones, R. C. (2004) Microwave or autoclave treatments destroy the infectivity of infectious bronchitis virus and avian pneumovirus but allow detection by reverse transcriptase-polymerase chain reaction. Avian Pathol. 33, 303–306.

    Article  CAS  PubMed  Google Scholar 

  10. Crookshanks, M., Emmersen, J., Welinder, K. G., and Nielsen, K. L. (2001) The potato tuber transcriptome: analysis of 6077 expressed sequence tags. FEBS Lett. 506, 123–126.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2008 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Nielsen, K.L. (2008). \(^\prime\)- and 3\(^\prime\)-RACE from LongSAGE Tags. In: Nielsen, K.L. (eds) Serial Analysis of Gene Expression (SAGE). Methods in Molecular Biology™, vol 387. Humana Press. https://doi.org/10.1007/978-1-59745-454-4_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-454-4_8

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-676-4

  • Online ISBN: 978-1-59745-454-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics