Matt:LabNotes/2014-1-17
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Testing non-specific binding of Shifted FISSEQ[edit]
Procedure[edit]
Using negative controls (E7 & F7) from Matt:LabNotes/2014-1-15
- Strip with 80% formamide for 10min @50C
- Rinse twice with H2O
- Image 488 channel
- Add 50ul 2uM Shifted FISSEQ preheated to 60C (1ul 200uM Dye + 30ul 100% formamide + 10ul 20X SSC + 59ul H2O)
- Incubate RT for 15min
- Wash 2 times with 2X SSC
- Image 488 channel
Results[edit]
- More signals from non-specific binding than from actual experiment before
- Since this is a negative control well it actually is a good sign in a way...
- Continued checking for non-specific binding here: Matt:LabNotes/2014-1-27#Testing_non-specific_binding_of_Shifted_FISSEQ