Skip to main content

Single-Cell RT-PCR cDNA Subtraction

  • Protocol

Part of the book series: METHODS IN MOLECULAR BIOLOGY™ ((MIMB,volume 461))

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

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   219.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   219.99
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

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. St. John TP, Davis RW (1979) Isolation of galactose-inducible DNA sequences fromSachromyces cerevisiaeMby differential plaque filter hybridisation. Cell 16:443.

    Article  CAS  PubMed  Google Scholar 

  2. Zimmerman CR, Orr WC, Leclerc RF, Barnard EC, Timberlake WE (1980) Molecular cloning and selection of genes regulated inAspergillusdevelopment. Cell 11:709.

    Article  Google Scholar 

  3. Timberlake WE (1980) Developmental gene regulation inAspergillus nidulans. Dev Biol 78:497.

    Article  CAS  PubMed  Google Scholar 

  4. Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N (1985) Enzymatic amplification of α-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anaemia. Science 239:1350.

    Article  Google Scholar 

  5. Rappolee DA, Wang A, Mark D, Werb Z (1989) Novel method for studying mRNA phenotypes in single or small numbers of cells. J. Cell Biochem. 39:1–11.

    Article  CAS  PubMed  Google Scholar 

  6. Brady G, Billia F, Knox J, Hoang T, Kirsch IR, Voura EB, Hawley RG, Cumming R, Buchwald M, Siminovitch K, Miyamoto N, Boehmelt G, Iscove N (1995) Analysis of gene expression in a complex differentiation hierarchy by global amplification of cDNA from single cells. Curr Biol 5:909–922.

    Article  Google Scholar 

  7. Brady G, Barbara M, Iscove NN (1990) Representative in vitro cDNA amplification from individual hemopoietic cells and colonies. Meth Mol Cell Biol 2:17–25.

    CAS  Google Scholar 

  8. Brady G, Iscove NN (1993) Amplified representative cDNA libraries from single cells. In: Wassarman PM, DePamphilis ML (eds.) Methods in enzymology, vol. 225. Academic, San Diego, pp. 611–623.

    Google Scholar 

  9. Trumper LH, Brady G, Bagg A, Gray D, Loke SL, Griesser H, Wagman, R, Braziel R, Gascoyne RD, Vicini, S. (1993) Single-cell analysis of Hodgkin and Reed-Sternberg cells: molecular heterogeneity of gene expression and p53 mutations. Blood 81:3097–3115.

    CAS  PubMed  Google Scholar 

  10. Dulac C, Axel R (1995) A novel family of genes encoding putative pheromone receptors in mammals. Cell 83:195–206.

    Article  CAS  PubMed  Google Scholar 

  11. Cumano A, Paige CJ, Iscove NN, Brady G (1992) Bipotential precursors of B cells and macrophages in murine fetal liver. Nature 356:612–615.

    Article  CAS  PubMed  Google Scholar 

  12. Cano-Gauci DF, Lualdi JC, Ouellette AJ, Brady G, Iscove NN, Buick RN (1993) In vitro cDNA amplification from individual intestinal crypts: a novel approach to the study of differential gene expression along the cryptvillus axis. Exp Cell Res 208:344–349.

    Article  CAS  PubMed  Google Scholar 

  13. Barr FG, Emmanuel BS (1990) Application of a subtraction hybridization technique involving photoactivatable biotin and organic extraction to solution hybridization analysis of genomic DNA. Anal Biochem.186:369–373.

    Article  CAS  PubMed  Google Scholar 

  14. Sugai M, Kondo S, Shimizu A, Honjo T (1998) Isolation of differentially expressed genes upon immunoglobulin class switching by a subtractive hybridization method using uracil DNA glycosylase. Nucleic Acids Res 26:911–918.

    Article  CAS  PubMed  Google Scholar 

  15. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

Download references

Acknowledgments

This work was supported by The Leukaemia Research Fund, The Cancer Research Campaign, and The Mark Richardson Memorial Trust. We thank David Masters for contributing to the latter part of this work.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2008 Springer Science + Business Media, LLC

About this protocol

Cite this protocol

Sakhinia, E., Weaver, D.L., Núñez, C., Brunet, C., Bostock, V., Brady, G. (2008). Single-Cell RT-PCR cDNA Subtraction. In: Sharpe, P.T., Mason, I. (eds) Molecular Embryology. METHODS IN MOLECULAR BIOLOGY™, vol 461. Humana Press. https://doi.org/10.1007/978-1-60327-483-8_44

Download citation

  • DOI: https://doi.org/10.1007/978-1-60327-483-8_44

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-431-9

  • Online ISBN: 978-1-60327-483-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics