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How To Please Ron/Brain

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miRNA Sensors (Theory)

  •  Troubleshoot
    •  Why didn't the oligos nanodrop??
    •  Why didn't the primers bind to LS_miR_144?

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miRNA Sensors (Lab Work)

  •  Golden Gate
    •  Low Sensor miR-144-miR-181c
  •  Transform
    •  High Sensor miR-30d
    •  High Sensor miR-146a
    •  High Sensor let-7f
    •  High Sensor miR-125b
    •  Low Sensor miR-144
    •  Low Sensor miR-181c
  •  Pick colonies to miniprep and midiprep
    •  pEXPR: MAV1212-TRE-L7ae (LR)
    •  pEXPR: MAV1212-hEF1a-GAL4VP16 (LR)
    •  pEXPR: MAV1212-TRE-GAL4VP16 (LR)
  •  Prepare and run a gel (Hyperladder 2 and 4% agaros gel w/ annealed target sites)
    •  High Sensor miR-30d TS
    •  High Sensor miR-146a TS
    •  High Sensor let-7f TS
    •  High Sensor miR-125b TS
    •  Low Sensor miR-144 TS
    •  Low Sensor miR-181c TS
  •  FACS cell prep
  •  Transfect L7ae Experiment
  •  Analyze FACS data

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NOTES

We have a new primer to sequence with (again, from Jeremy)

We now have FACS DATA