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Total RNA Extraction from Rhizobium etli Pellets

Instrument: Precellys® 24

Sample: Rhizobium etli cell pellets or bacteroid pellets

Protocol:

  • Add R. etli cell pellets (from 20 - 40 ml of bacteria culture) or bacteroid pellets (prepared from Phaseolus vulgaris root nodules) to the 2 ml microorganism lysing kit.
  • Add 1 ml TRIzol Plus RNA Purification kit (Invitrogen).
  • Homogenize in the Precellys at 6500 rpm, 2 x 45 seconds with a 30 second pause.

Results:

Samples were analyzed for RNA quality using Experion RNA StdSens Chips (see figure below) and all samples had an RNA quality indicator of 10. Additionally, RNA quantity and purity was assessed using the NanoDrop ND-1000, and both the A260/A280 ratio and A260/A230 ratio for all samples were ≥ 2.

Graph demonstrating the high quality total RNA obtained from R. etli using the Precellys 24, illustrating the fragmentation 23S rRNA.

Graph demonstrating the high quality total RNA obtained from R. etli using the Precellys 24, illustrating the fragmentation 23S rRNA.  (a) small RNA peak including 5S rRNA and the ncRNAs (b) 23S fragment of ~135 bp (c) two 23S fragments of ~1300 bp (d) 16S rRNA (e) intact 23S rRNA.