Plant Salt Tolerance pp 335-350

Part of the Methods in Molecular Biology book series (MIMB, volume 913) | Cite as

Transcriptomics on Small Samples

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

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.

Key words

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

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Stuart J. Roy
    • 1
  • Simon J. Conn
    • 2
  • Gwenda M. Mayo
    • 1
  • Asmini Athman
    • 3
  • Matthew Gilliham
    • 3
  1. 1.Australian Centre for Plant Functional Genomics and School of AgricultureFood and Wine & Waite Research InstituteGlen OsmondAustralia
  2. 2.European Molecular Biology LaboratoryGrenoble OutstationGrenobleFrance
  3. 3.School of Agriculture, Food and Wine, & Waite Research InstituteUniversity of AdelaideGlen OsmondAustralia

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