RNA Extraction from Caenorhabditis
Instrument: Precellys® 24 Dual
Sample: C.elegans from a full 9cm petri dish (either mixed stage, or staged, depending on requirements)
Protocol:
- Precellys® lysing kit: 03961-1-004 (0.5mm glass beads)
- Add sample: Suspension of C.elegans gently pelleted by centrifugation (1500rpm for 1min) and washed three times with M9 buffer before resuspending in 500μl of M9 buffer.
- Run the Precellys at 6400 rpm, 2x10 sec, repeat for a total time of 40secs.
Results:
RNA was isolated directly from the extract using Qiagen RNeasy kit before being checked for quality and quantity using NanoDrop analysis.
On average a yield of 1.5 – 2 μg/μl of RNA was obtained per 9cm petri plate. This RNA was then used for RT-PCR to quantify gene expression studies as shown
