Riboprobe purification question

Michael L. Sullivan mlsulliv at facstaff.wisc.edu
Mon Jan 14 09:28:19 EST 2002

I've found that the spin columns used for cleaning up sequencing reactions
(e.g. Autoseq G-50 from Pharmacia, no affiliation) work nicely for this


>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

Michael L. Sullivan, Ph.D

U.S. Dairy Forage Research Center
1925 Linden Drive West
Madison WI, 53706

(608) 264-5144 Phone
(608) 264-5147 Fax


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