User:Karmella Haynes/Notebook/Polycomb project/2010/08/25

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08/25/10

  • ✓ Microscopy: fix and block slides



Microscopy
> Check for RFP expression: punctate spots not as clear as before in 130-4 (used half as much dox for induction this time)
> Dissolve paraformaldehyde in 1xPBS (dissolve with occasional shaking at 60°C, takes ~2 hrs.) to make 4% paraformaldehyde
; cool to room temp
--> Formaldehyde solution contains some methanol which might cause permeabilization too soon
> Permeabilize cells with 0.5% Triton X-100
> Rinse 2x w/ 1xPBS
> Block w/ 5% normal horse serum @ 4°C/ overnight



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