Riboprobe purification question

Magdalena Popesco mpopesco at columbus.rr.com
Sun Jan 13 13:53:40 EST 2002


I am labeling a riboprobe (1.7kb) with 35S with the in vitro transcription
kit from Promega. The purification method
used, following the in vitro transcription reaction, was standard phenol
extraction and ethanol precipitation.  This method has been very
inconsistent between experiments. I am not content with the yield recovery.

I have a question regarding the preparation/purification of riboprobes.
What purification method can be used following
the in vitro transcription reaction to remove unincorporated
35S-radionuleotides?  Is there a particular column or kit
that can be used?  Is there a reference you may direct me for riboprobe
labeling and purification?

Thanks in advance,

popesco.1 at osu.edu

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