Sam’s Notebook: RNA Isolation and Quantification – C.bairdi Hemolymph Pellets in RNAlater

TL;DR – Recovered absolutely no RNA from any sample! However, I did recover DNA from each sample.

Isolated RNA from the following 23 hemolymph pellet samples:

  • 6128_112_9
  • 6204_114_9
  • 6141_123_9
  • 6245_126_9
  • 6240_134_9
  • 6260_136_9
  • 6257_138_9
  • 6259_139_9
  • 6258_140_9
  • 6255_143_9
  • 6256_146_9
  • 6265_155_9
  • 6266_156_9
  • 6261_164_9
  • 6120_165_9
  • 6251_167_9
  • 6262_168_9
  • 6243_173_9
  • 6263_179_9
  • 6264_180_9
  • 6200_208_12
  • 6204_252_12
  • 6190_256_12

Isolated RNA using the Quick DNA/RNA Microprep Kit (ZymoResearch; PDF) according to the manufacturer’s protocol for liquids/cells in RNAlater.

  • Used 35uL from each RNAlater/hemocyte slurry.
  • Mixed with equal volume of H2O (35uL).
  • Retained DNA on the Zymo-Spin IC-XM columns for isolation after RNA isolation.
  • Performed on-column DNase step.
  • RNA was eluted in 15uL H2O

RNA was quantified on the Roberts Lab Qubit 3.0 using the RNA High Sensitivity Assay (Invitrogen), using 2uL of each sample.