Unravelling Single Cell Genomics
Chapter 9: Gene Analysis of Single Cells
Published:18 Oct 2010
Special Collection: 2010 ebook collection , 2010 ebook collection , 2010 materials and nanoscience subject collectionSeries: Nanoscience & Nanotechnology
Bruno Cauli, Bertrand Lambolez, 2010. "Gene Analysis of Single Cells", Unravelling Single Cell Genomics, Jean-Christophe Baret, Bruno Cauli, Max Chabert, Valerie Abecassis-Taly, Petra Dittrich, Emmanuel Fort, Christoph Klein, Joel Lachuer, Bertrand Lambolez, Nicholas Le Novere, Severine le Gac, Laili Mahmoudian, Yann Marcy, Bernhard Polzer, Joelle Vinh, Tania Vitalis, Nathalie Bontoux, Marie-Claude Potier, Luce Dauphinot, Harold Craighead, Harry Kroto, Paul O'Brien, Royal Society of Chemistry
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Shortly after the initial demonstration of the feasibility of gene analysis at the single cell level,1 the two major technical trends enabling gene expression analyses of single cells developed in the early 90s. These developments were aimed either at whole transcriptome analysis based on RNA amplification,2,3 or at limited gene expression profiling using RT-PCR for correlating molecular and functional properties.4 The brain complexity and cellular diversity has been a strong incentive for the development of these tools at a time when many of the major constituents of neurotransmission had been cloned. Both techniques initially relied on the use of the patch-clamp technique5 to harvest selectively the cell's mRNAs. In this chapter we will detail the key steps, which assessed the reliability and functional relevance of the “single cell RT-PCR after patch-clamp” technique (scPCR, Lambolez et al., 1922), and describe its evolutions. We will also share our observations on the design and interpretation of scPCR experiments and discuss the limits of this approach.