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13.2.20

Objectives

  • Make 1XTAE from 50X stock solution
  • Run gel of VC3113 PCR and visualize
  • Order HpyCH4IV for PR671 diagnostic
  • Put PR671 PCR product into freezer for use when enzyme cutter arrives
  • Experiment with OP50 lawns
  • Starve GRU101
  • Record GRU101 with OP50
  • Set up OP50-GFP ON
  • Maintain strains
  • Update video log

Observations

  • 1XTAE - 20ml 50XTAE in 1L dH20
  • 100ml gel at 1.5% (1.5g agarose) with 8ul ethidium bromide (0.5ul/ml final concentration)
    • Gel loaded L A1 A2 … A8 L
    • entire 20ul of PCR product used
    • 10ul for each ladder
    • A1 faint - was using the wrong pipette to load it so either not all of PCR product made it in there or was maybe more diffuse
  • GRU101
    • 0032 - G5, “dab” lawn using soft pipette tip, 10 worms
      • had 4 worms going inside lawn even 0.5cm on either side and with such a tiny lawn
      • Monika suggests to do this with OP50-GFP but just crank up the exposure/gain for the photo - touch the tip to the agar by hand
    • 0033 - G0 2ul, lawn using soft pipette tip, 10 worms
  • Will need to redo gel tomorrow - Tm probably too high - try 52C

Gel

TOMORROW

  • Be sure to maintain plates 1-10 of VC3113 F3 until genotyping happens!!!!
  • Maintain strains
  • Starve GRU101
  • Seed “dab” lawns
  • Record GRU101 response to “dab” lawns and hopefully get longer trajectories
wiki/documentation/wiki/documentation/nicolina/13.2.20.1581612802.txt.gz · Last modified: 2020/02/13 08:53 by nzjacic

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