Transcriptomics on small samples

Stuart J. Roy, Simon J. Conn, Gwenda M. Mayo, Asmini Athman, Matthew Gilliham

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

4 Citations (Scopus)

Abstract

Interrogating the cell-specific transcriptome forms an important component of understanding the role that specific cells play in assisting a plant to overcome abiotic stress. Among the challenges arising when extracting RNA from individual plant cells are: the isolation of pure cell populations; the small yield of material when isolating specific cell types, and ensuring an accurate representation of the transcriptome from each cell type after amplification of RNA. Here we describe two approaches for isolating RNA from specific cell types - single cell sampling and analysis (SiCSA) and laser capture microdissection. Isolated RNA can then be directly sampled qualitatively using reverse transcription PCR (RT-PCR) or amplified for profiling multiple specific genes using quantitative RT-PCR and genome-wide transcript analyses.

Original languageEnglish
Title of host publicationPlant Salt Tolerance
Subtitle of host publicationMethods and Protocols
PublisherHumana Press
Pages335-350
Number of pages16
ISBN (Print)9781617799853
DOIs
Publication statusPublished - 1 Jan 2012
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume913
ISSN (Print)1064-3745

Keywords

  • Laser capture microdissection
  • RNA isolation
  • Single cell sampling and analysis (SiCSA)
  • Transcriptome

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