Preparation of Total RNA from Monosiga brevicollis and Other Choanoflagellates
- 1Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94704, USA
- 2Department of Integrative Biology, University of California, Berkeley, Berkeley, CA 94704, USA
- 3Canadian Institute for Advanced Research, Toronto, Ontario M5G 1Z8, Canada
- 4Department of Biology, University of York, York YO10 5YW, United Kingdom
- 5School of Biosciences, University of Birmingham, Birmingham B15 2TT, United Kingdom
- ↵6Corresponding author (b.s.c.leadbeater{at}bham.ac.uk)
INTRODUCTION
Choanoflagellates are heterotrophic nanoflagellates: small, colorless protozoa that are present in marine and freshwater environments as well as in hydrated soils. Because they are the closest living relatives of the metazoa, the study of their cell biology and genomes promises to provide new insights into metazoan ancestry and origins. The preparation of RNA is an important prerequisite for characterizing the gene expression profile of choanoflagellates under different conditions. Although most standard protocols (e.g., those using TRIzol) are sufficient for many purposes, the approach described here minimizes RNA degradation and provides a high-quality template for downstream protocols, including real-time polymerase chain reaction.
