Grace’s Notebook: Tri Reagent Extraction_protocol

Tri-reagent extraction protocol

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

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