Hosuk:LabNotes/2013-5-1: Difference between revisions
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====Debugging plan (re-organized)==== | ====Debugging plan (re-organized)==== | ||
*Re-planning from the previous debugging plan ([[Hosuk:LabNotes/2013-4-1|04/01]]) | |||
=====1. mRNA population check after fixing===== | =====1. mRNA population check after fixing===== | ||
*[[Hosuk:LabNotes/2013- | *mRNA were observed from the previous result([[Hosuk:LabNotes/2013-3-17|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([[Hosuk:LabNotes/2013-3-17|03/17]]), but just do again with the new samples fixed at 05/01 |
Revision as of 21:00, 2 May 2013
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