RNA Fragmentation – Rick’s trout RBC samples prepped earlier today (see below)

Samples were fragmented according to the Whole Transcriptome Kit protocol. Samples were then cleaned up using Invitrogen’s Modified RiboMinus Concentration Module, according to protocol. Briefly:

  • Added 1X volume of binding buffer (100uL)
  • Added equal volume of 100% EtOH so that [EtOH] = 50% (200uL)

Followed remainder of protocol and eluted with 20uL of H2O. Samples were spec’d.

Results:

Assuming the NanoDrop readings are accurate (according to the Whole Transcriptome Kit, these may NOT be accurate), got yields of ~93ng for the RBC Control sample and ~132ng for the RBC poly1:C sample. Transferred 1.5uL of each sample to separate 0.5mL tubes for submission to the Bioanalyzer. These were stored @ -80C in the “Bioanalyzer Samples” box. The remainder of the samples were stored @ -80C in the “Samples from Other Researchers” box.

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