insitu hybridization

Robert Brown rcbrown at casbah.acns.nwu.edu
Thu Dec 19 15:11:44 EST 1991


Hello mobio jocks,

I got a little problem here with in-situ hybridization. I am currently using 
S-35 RNA probes. I have gotten the procedure to work with chicken brains. 
However, pituitary seems to be another matter. Sense control probes gave a 
signal that was comparable to antisense probes. I am interested in conditions 
that would relate to pituitary or other tissues. Specifically, cpms/ml, probe 
size, specific activity of the probes, stringency of the washes (do you use 
mercaptoethanol in your washes), temperature of washes, how long you can store
tissue in OTC at -70 degrees. I really need this procedure to work because I 
am using it to screen PCR clones. I would appreciate any help I can get.


Thanks,
Clark Brown

 



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