Expression analysis of rare cellular subsets: direct RT-PCR on limited cell numbers obtained by FACS or soft agar assays
Victor Ho, Shi Yun Yeo, Kamini Kunasegaran, Duvini De Silva, Gerard A Tarulli, P Mathijs Voorhoeve, Alexandra M Pietersen
BIOTECHNIQUES | BIOTECHNIQUES OFFICE | Published : 2013
Since tissues and tumors are heterogenous populations containing different cell types, their transcriptomes are blends of multiple mRNA expression profiles. Although fluorescence-activated cell sorting (FACS) allows isolation of individual cell types, RNA isolation and quantification remain problematic from rare subsets, such as tissue stem cells. Likewise, identification of transcriptional changes relevant to the tumorigenic potential of mammalian cells while they are actively growing as colonies in soft agar is also hampered by limited amounts of starting material. Here we describe a convenient method that fills the gap between single cell and whole tissue mRNA analysis, enabling mRNA quan..View full abstract
Awarded by Singapore Ministry of Health's National Medical Research Council
Awarded by Individual Research Grant
We would like to thank Manu Beillard for valuable input. This research was supported by the Singapore Ministry of Health's National Medical Research Council under its Exploratory Development Grant (NMRC/EDG/0066/2009) and Individual Research Grant (NMRC/GMS/1250/2010) scheme.