Daniel:Notebook/RNAFISH/2014-10-10
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ABP Trial Run[edit]
This is a trial run of the Allen Brain Project RNAFISH. I will be using RBFOX3 and FOXP2 as probes, since RBFOX3 is non-specific and FOXP2 is layer specific, giving me one general signal and one localized signal as comparisons.
Tissue Fixation[edit]
- Fix slides/sections in freshly prepared 4% paraformaldehyde/PBS depc, pH 7.2-7.4 at room temp. for 30 minutes
- Wash 2x in 2xSSPE for 5 minutes each
- Permeabilize in 0.6% Triton X-100/2xSSPEdepc for 30 minutes, room temp
- Wash 3 x 5 minutes in 2xSSPE
- Treat slides for 10 minutes/room temp in acetylation buffer
- Make the buffer just before use
- Wash 3x 5 mins 2xSSPE
- Dehydrate in 30, 50, 70, 100, 100 ethanols 2 mins ea.
- Make solutions from absolute ethanol and depc/H20*.
- Dry in oven at 50-65C.