Why DTT in in situ hybridization???

Istvan Barna barna at koki.hu
Thu Oct 21 03:31:27 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. 

OK, this is absolute clear...

But I use DIG-labelled riboprobes!! 
Do you think there is any protective effect of DTT on this system, too.

Steve



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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