Transcriptomics on small samples.
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.
Australian Centre for Plant Functional Genomics and School of Agriculture, Food and Wine & Waite Research Institute, Glen Osmond, SA, Australia. stuart.roy@acpfg.com.au
Stuart J
SJ
Roy
Simon J
SJ
Conn
Gwenda M
GM
Mayo
Asmini
A
Athman
Matthew
M
Gilliham
D017360:Arabidopsis / Q000235:genetics; D020869:Gene Expression Profiling / Q000295:instrumentation / Q000379:methods*; D061346:Laser Capture Microdissection / Q000295:instrumentation / Q000379:methods; D018517:Plant Roots / Q000235:genetics; D010944:Plants / Q000235:genetics*; D018749:RNA, Plant / Q000302:isolation & purification; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods; D059467:Transcriptome
D018749:RNA, Plant