Biomod/2011/TUM/TNT/LabbookA/2011/10/10

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10th Oct 2011

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Contents

TEM Measurements of BM_2 with EtBr

Sample Preparation

Sample: Biomod_2_B2, Biomod_2_B3, Biomod_2_B4

  • In equilibrium each 7 base pairs one EtBr molecule should have bound to the structure
  • All dilutions are made with FOB20
  • cBM_2 ≈ 0.25 nM, VBM_2 (@ ≈ 10 nM) = 1 µl
  • cEtBr = 2.27 µM, VEtBr (@ 5 µM) = 18.6 µl
  • VFOB20 = 20.9 µl
  • Vtot = 40 µl
  • Vortexed sample for 10 sec and shortly centrifuged it
  • Incubate for 5 min before making grids

Sample: Biomod_2_B5, Biomod_2_C1, Biomod_2_C2

  • In equilibrium each 21 base pairs one EtBr molecule should have bound to the structure
  • All dilutions are made with FOB20
  • cBM_2 ≈ 0.25 nM, VBM_2 (@ ≈ 10 nM) = 1 µl
  • cEtBr = 0.69 µM, VEtBr (@ 5 µM) = 5.5 µl
  • VFOB20 = 33.5 µl
  • Vtot = 40 µl
  • vortexed sample for 10 sec and shortly centrifuged it
  • Incubate for 5 min before making grids

Sample: Biomod_2_C3, Biomod_2_C4, Biomod_2_C5

  • Controls without any DNA binder
  • cBM_2 ≈ 0.25 nM, VBM_2 (@ ≈ 10 nM) = 1 µl
  • VFOB20 = 39 µl
  • Vtot = 40 µl
  • Vortexed sample for 10 sec and shortly centrifuged it
  • Incubate for 5 min before making grids

Experiment

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