Matt:LabNotes/2014-3-19
Jump to navigation
Jump to search
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
- Pre-heat 10uM MALAT1_Template in 2X SSC to 85C (50ul 100uM MALAT1_Template + 50ul 20X SSC + 400ul H2O)
- Add 50ul to each well
- Incubate @RT for 15min
- Aspirate but don't wash
- Add 50ul BS(PEG)9 mix (10ul BS(PEG)9 + 490ul 1X PBS)
- Incubate @RT for 1hr
- Wash with 1X PBS twice
- Add 50ul 1M Tris pH 8.0
- Incubate @ RT for 30min
- Wash with 1X PBS twice