lac Z embryo staining

kposey kposey at uh.edu
Thu Feb 10 15:48:33 EST 2000


Dear  Newsgroup:

I am trying to stain embryos using X gals staining buffer. The
proceedure is outlined below.  I am getting good staining but the
embryos appear dehydrated.  PLEASE HELP!

-dechorinate 2 min in 50% bleach
-Fix in 2 ml heptane and 2 mix fix (3.7% formaldehyde in PBS)
    vortex 5 sec incubate 5 min repeat total 3 X
-most of Heptane is removed and mixture is moved to watch glass to let
heptane evaporate (I have also tried not doing the drying and remove
heptane as much as I can and remove fix and then add PBST)
-remove fix
-add PBST
-wash 3-5X in PBST
-add X gal staining buffer with freshly diluted X gal
-incubate 37 deg wrapped in foil until color develops.

Thanks

Karen Posey
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