Grace’s Notebook: October Goals

Crab Project

  • Identify successful RNA extraction protocol
  • Get extracting RNA going so that we can plan to send some more samples for library prep and sequencing
  • Publish more DecaPod episodes (about 2 – one every two weeks)

2015 Oysterseed Project

  • Work through what Emma sent to see that I understand what she did
  • Start analyzing the data

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Grace’s Notebook: Extract RNA from non-lyophilized C. bairdi hemolymph

Today I am extracting RNA from four practice samples using Tri-reagent, and without lyophilizing (lyophilizer is out of commission again for now).

Protocol (from Tri-reagent protocol in Sam’s notebook)

  1. Added 1mL TriReagent to each tube
  2. vortexed to mix/dissolve solute
  3. incubated 5mins at RT
  4. added 200uL of chloroform, vortexed 15s to mix
  5. incubated at RT for 5mins
  6. centrifuged 15mins, 12,000g, 4˚C
  7. transferred aqueous phase to new tube
  8. added 500uL isopropanol to aqueous phase
  9. mixed, incubated at RT for 10mins
  10. centrifuged 8mins, 12,000g, at RT
  11. discarded supernatant
  12. added 1mL 75% ethanol
  13. centrifuged 5mins, 12,000g at RT
  14. discarded supernatant
  15. resuspended in 10uL of 0.1% DEPC-treated H2O.

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