RNA Isolation and Quantification - C.bairdi RNA from Hemolymph Pellets in RNAlater

Based on qPCR results testing for residual gDNA from 20200225, a set of 24 samples were identified that required DNase treatment and/or additional RNA. I opted to just isolate more RNA from all samples, since the kit includes a DNase step and avoids diluting the existing RNA using the Turbo DNA-free Kit that we usully use. 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.


Qubit results (Google Sheet):

Most samples have good yields. Only a few are a bit low. Will check all samples for residual gDNA.