Fog in In Vitro Translations
mmorales at acpub.duke.edu
Sat Mar 23 13:55:19 EST 1996
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,
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