RNA extraction from a single worm (ideally L1)

Model Organism: Worms Methods & Reagents
1

Greetings,

We are interested in extracting RNA from a SINGLE, early development worm (preferably L1). Our scientific interest is in random variation in genome-wide expression between individual genetically identical worms. So far we have yet to find a protocol that reliably allows us to do this, although the source of variability may be in the quantification rather than in the extraction per se.

I have two questions:

  1. Does anyone have a reliable method of extracting RNA (and then determining RNA quantity and quality) from a single, young worm? Our greatest successes so far have included freeze drying and bead beating followed by extraction using Ambion RNAqueous-micro reagents and columns. We have had limited success with Trizol extractions, possibly because the chemical cuticle disruption is not efficient with such low inputs?

  2. Does anyone have suggestions for methods of picking a single L1 worm that would limit bacterial contamination in downstream nucleic acid preparations?

Any suggestions would be greatly appreciated.

Thanks very much,

-Charlie Baer