Matt:LabNotes/2014-3-19

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Revision as of 00:52, 19 March 2014 by >Mzcai (Created page with "==Padlock Probe Concentration Test using Fixed Template== *96-well plate NCI-H1975 Cells **Alan already marked locations with good cell density *Test concentrations NegCtrl, ...")
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Padlock Probe Concentration Test using Fixed Template

  • 96-well plate NCI-H1975 Cells
    • Alan already marked locations with good cell density
  • Test concentrations NegCtrl, 10pM, 50pM, 100pM, 10nM
    • 2 wells for each


Fixing Template in cells

  1. Pre-heat 10uM MALAT1_Template in 2X SSC to 85C (50ul 100uM MALAT1_Template + 50ul 20X SSC + 400ul H2O)
  2. Add 50ul to each well
    • Incubate @RT for 15min
  3. Aspirate but don't wash
  4. Add 50ul BS(PEG)9 mix (10ul BS(PEG)9 + 490ul 1X PBS)
    • Incubate @RT for 1hr
  5. Wash with 1X PBS twice
  6. Add 50ul 1M Tris pH 8.0
    • Incubate @ RT for 30min
  7. Wash with 1X PBS twice