How To Please Ron/Brain

miRNA Sensors (Theory)

  • Alternative backbones? Check iGEM 2013
  • Actual list of experiments
    • TAL14 & GAL4 (to test promoter)
    • MAV1212-hEF1a-eBFP2
  • Make/Find primers
    • To sequence both low sensors

miRNA Sensors (Lab Work)

  • Sequence/digest plasmids to verify - may be unnecessary. Emailed Brian.
    • EF
    • GH - don't have this
    • pEXPR: MAV1212-Hef11a-eBFP2
    • Mutant Hef1a-2xKturn-eGFP
    • Wild Type Hef1a-2xKturn-eGFP
  • Pick colonies and grow up
    • MAV1212-mRFP
    • L1_TALER14_L2
    • pTAL14-mKate-pEST
    • MAV1212-Hef1a-eBFP2 for midiprep (pending Brian's approval)
    • EF low sensor for midiprep (pending Brian's approval)
    • Mutant Hef1a-2xKturn-eGFP for midiprep (pending Brian's approval)
    • WT Hef1a-2xKturn-eGFP for midiprep (pending Brian's approval)
  • Miniprep
    • GH - don't have this
  • Transformations
    • GH (Golden Gate)
    • Hef1a-L7ae (LR)
    • Hef1a-TAL14 (LR)
  • BP
    • P4-donor-P1R with TAL14-mKate pEST

NOTES

BP rxn didn't yield colonies --> re-doing it and letting it grow overnight this time

GH plate has no colonies :/

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