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Detecting Gene Expression in Lymphoid Microenvironments by Laser Microdissection and Quantitative RT-PCR

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Germinal Centers

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1623))

Abstract

Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) is a valuable tool for measuring gene expression in cells and tissues. Unique challenges are encountered when studies are performed on cells microdissected from small specific areas of frozen animal or human tissue. This chapter describes the analysis of gene expression of chemokines and cytokines that are important for the differentiation and migration of germinal center (GC) derived plasmablasts/plasma cells and memory B cells by using laser capture microdissection (LCM) and qRT-PCR to examine tissue sections.

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References

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Acknowledgment

This work was supported by grants from the MRC (Topjabs, G1001390) and BBSRC (BB/M025292/1), and received funding from the People Programme (Marie Curie Actions) of the European Union’s Seventh Framework Programme FP7/2007-2013 under Research Executive Agency grant agreement № 315902. LGI gratefully acknowledges receipt of a Marie Curie Research Associate post. GB and K-MT are partners within the Marie Curie Initial Training Network DECIDE (Decision-making within cells and differentiation entity therapies).

Competing Financial Interests

The authors declare no competing financial interests.

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Authors and Affiliations

  1. Institute for Immunology and Immunotherapy, Institute for Biomedical Research, University of Birmingham Medical School, Vincent Drive, B15 2TT, Birmingham, UK

    Yang Zhang, Laura Garcia-Ibanez, Geoffrey Brown & Kai-Michael Toellner

Authors
  1. Yang Zhang
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  2. Laura Garcia-Ibanez
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  3. Geoffrey Brown
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  4. Kai-Michael Toellner
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Corresponding authors

Correspondence to Yang Zhang or Kai-Michael Toellner .

Editor information

Editors and Affiliations

  1. Immunity and Cancer Laboratory, The Francis Crick Institute, and King’s College London, London, United Kingdom

    Dinis Pedro Calado

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Zhang, Y., Garcia-Ibanez, L., Brown, G., Toellner, KM. (2017). Detecting Gene Expression in Lymphoid Microenvironments by Laser Microdissection and Quantitative RT-PCR. In: Calado, D. (eds) Germinal Centers. Methods in Molecular Biology, vol 1623. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7095-7_2

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  • DOI: https://doi.org/10.1007/978-1-4939-7095-7_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7094-0

  • Online ISBN: 978-1-4939-7095-7

  • eBook Packages: Springer Protocols

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