35S-labelled oligo purification
Dr. Duncan Clark
Duncan at nospam.demon.co.uk
Wed Feb 2 07:51:14 EST 2000
In article <8791g0$2mj$1 at beta.qmw.ac.uk>, News
<D.P.McLaughlin at mds.qmw.ac.uk> writes
>The oligos are HPSF pure synthesised by MWG biotech - no problems in the
>past and EtOH
>precipitation works well - just looking for a quicker method. Pharmacia
>assure us that the unincorporated 35SdATP will wash off the column, but that
>interacton with the 35S labelled oligo is 'ionic'. Trouble is, we can wash
>unincorporated 35S after a few washes, but the labelled oligo sticks until
>vigorously and then eluted. It is all very puzzling since the product
>guides for all
>their microspin columns suggests G25 for labelled oligos
Am I missing something here? How are you labelling oligos with 35S dATP?
Are you not kinasing using T4PNK and gamma labelled 35S ATP?
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....
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