Fog in In Vitro Translations

[Michael Morales]

John (JPFELL00 at ukcc.uky.edu) wrote:
:  
: We are trying to translate protein from pBluescript derived RNA.  We have used
: both rabbit retic and wheat germ lysates, but get a fog of 35S label that goes
: from about 40 kD to the bottom of the gel.  Does anyone know how to prevent
: this or get rid of it?  thanks.
:  
1) It may be acylated tRNA.  RNAse treatment after translation should get
rid of it.  2) If you're using the Tris-Tricine (I think) buffer in your gel 
for imporved resolution of LMW proteins, you'll get an 35S "fog."  Go to a 
conventional SDS-PAGE with tris-glycine.

Hope this helps; good luck, 


Mike Morales