Hosuk:LabNotes/2013-5-1
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Debugging plan (re-organized)
- Re-planning from the previous debugging plan (04/01)
1. mRNA population check after fixing
- mRNA were observed from the previous result(03/17), but just do again with the new samples fixed at 05/01
- Detail procedure
- Fix cells and permeabilize using 70% ethanol as before
- Mix 5 ul fluorescent primer (Cy3-polydT(50 mer)) in 200 ul 2xSSC/10% dextran sulfate/10% formamide
- Add to cells and incubate at 37C for 1 hour
- Wash using 2xSSC/10% formamide for 15 min each twice
- Aspirate and add 2x SSC with DAPI or Hoescht dye for nuclear staining and then image
1. mRNA population check after fixing
- mRNA were observed from the previous result(03/17), but just do again with the new samples fixed at 05/01