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/2013-4-1|04/01]]
*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
    1. Fix cells and permeabilize using 70% ethanol as before
    2. Mix 5 ul fluorescent primer (Cy3-polydT(50 mer)) in 200 ul 2xSSC/10% dextran sulfate/10% formamide
    3. Add to cells and incubate at 37C for 1 hour
    4. Wash using 2xSSC/10% formamide for 15 min each twice
    5. 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