Post subject: In Situ HELP! Cannot get ANY signal at all!
I've been trying to set up in situ hybridization in our lab for several months now but have had no luck.
Every protocol i've tried has resulted in absolutely zero signal when I run my slides through our phospho imager.
I've tried my own probes, along with probes made by professionals and still no signal which leads me to believe that the way I'm running the procedure is not going right.
I have photos from my phospho imager if needed but there's basically nothing.
Fri Feb 11, 2011 7:57 pm
Joined: Oct 11, 2004
Post subject: "the procedure is not going right"
Dont think i can help you? Most have more ISH-experience than me.
If you tell us more about the probe(s) and post a detailed procedure im sure someone can assist you.
Wed Feb 23, 2011 4:02 pm
Joined: Mar 07, 2011
Do you have any way to confirm that your RNA of interest is expressed in the sample? Or is the experiment to see IF the RNA is expressed in the sample?
Have you confirmed that the RNA in the sample is intact? There are some good control probes that you can buy that will determine whether RNA is intact. Ubiquitin (UbC) is a good control.
That's where I would start.
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