dsDNA clean-up with Sephadex

Dennis J. Templeton djt2 at po.CWRU.Edu
Wed Nov 20 13:40:40 EST 1991


In a previous article, cbar89 at vaxa.strath.ac.uk () says:

>Does anyone out there have a suitable method for the removal of nucleotides
>and, if possible, 20 mer primers from a reaction mixture using Sephadex
>in a home made spin column. The dsDNA in the mixture is at least 120 bp and 
>requires some concentration as well as clean up.
>The sample should also be desalted in the clean up step.
>
>Any help as to column preparation, spin times and g-force would be greatly
>appreciated.
>
>
>Martin J Pearce
>Forensic Science Unit
>University of Strathclyde
>Glasgow UK.
>
>PS Any suggestions for alternative techniques are welcome. (But we haven't
>got a centrifuge that will take Centricon devices!!)
>

Amicon (I think) or maybe Schleicher and Schule makes a centricon-like device 
that is the size and shape of a microfuge tube....

This might be an option.

We use spin columns with p6-DG (from BioRad) frequently but I'm not sure 
you can separate 120 nt from primers or worse primer-dimers (~40 nt?)

good luck

dennis



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