We don't treat our slides because they come clean already (Fisher Superfrost
When our in situ's don't work, we just make all of the solutions over and
buy new NBT/BCIP and DIG antibody. This has always worked for us, though it
can get expensive!
You can check if your probe is working by dotting it on filter paper and
going through your protocol (obviously you can skip stuff like proteinase K
or fixation). This will also check all of your reagents.
Also, make sure the RNase Away is not left on your utensils, we do a short
rinse with DEPC water after treating them.
One other small thing--check your hybridization oven temperature with a
thermometer. Our digital reading has always been off by five degrees.
When you're riding in a time machine way far into the future, don't stick
your elbow out the window, or it'll turn into a fossil.
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