Hosuk:LabNotes/2013-10-19: Difference between revisions

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>Hosuki78
(Created page with "*LabNote ===Rolonies with fresh cells (96well plate)=== ====Process==== *Start (10/17) *End RCA : 1:30pm 10/19 *...")
 
>Hosuki78
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*End RCA : 1:30pm 10/19
*End RCA : 1:30pm 10/19
*...
*...
====Result====
*Not good : there are rolonies, but almost no ACTB
*Cells in 6 out of 8 wells don’t look good, they were detached, less cells in wells.. WHY? Something happened when fixing?
*Cells were aggregated or became chunk…  need to mix when split
*Fixing two step : 37% Formalin  10% Formalin
*The very first protocol was two step fixing, but Jonathan has been doing one step (10% Formalin), and in their manuscript as well.
*But… I want to try two step fixing and compare to one step fixing…

Revision as of 19:14, 20 October 2013


Rolonies with fresh cells (96well plate)

Process

  • Start (10/17)
  • End RCA : 1:30pm 10/19
  • ...


Result

  • Not good : there are rolonies, but almost no ACTB
  • Cells in 6 out of 8 wells don’t look good, they were detached, less cells in wells.. WHY? Something happened when fixing?
  • Cells were aggregated or became chunk…  need to mix when split
  • Fixing two step : 37% Formalin  10% Formalin
  • The very first protocol was two step fixing, but Jonathan has been doing one step (10% Formalin), and in their manuscript as well.
  • But… I want to try two step fixing and compare to one step fixing…