Why DTT in in situ hybridization???
George Flentke
gflentke at facstaff.wisc.edu
Wed Oct 20 05:14:13 EST 1999
In article <380C80EE.F668FD96 at koki.hu>, barna at koki.hu wrote:
>Maybe you are absolute right, but in situ hybridization solutions do not
>contain RNAsin :)))
>
>it contains:
>
>Formamide
>Dextran
>Nacl
>Denhard's
>TRIS
>EDTA
>tRNS
>DTT
>and the probe
>
Originally, all probes were S-35. The DTT helped keep the alpha-S from
oxidizing off the probe. Thus it was a stabilizer for the probe.
DTT does seem to be thought of as necessary for RNA'sin stability. The
problem with the descrepancy might be due to long term versus short term
protein stability. I always keep a bit there, just in case. With the binding
mechanism not well understood, it can hurt.
George
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