plastic disposable columns

tan at aeolus.vmsmail.ethz.ch tan at aeolus.vmsmail.ethz.ch
Wed Mar 16 11:15:41 EST 1994


In article <79712.a.hanly at cheque.uq.oz.au> "Allison Hanly"
<a.hanly at cheque.uq.oz.au> writes:
>Song Tan  <tan at aeolus.vmsmail.ethz.ch> wrote:

>>Slight tangent to this discussion, but has anyone successfully used these
>>BioRad biospin columns with plasmid DNA?  I use Sepharose CL-4B or S400 spun
>>columns as the last step in my plasmid minipreps but when I tried using these
>>resins in the BioRad biospin columns, I got no DNA back in the spun column
>>"eluent".  Recovery from the home made spun columns (siliconized glass wool to
>>plug a Gilson blue tip) was fine, so I attributed the loss to the biospin
>>columns.  Would be nice to hear others' experiences.
>>
>What rotor type did you use?
>Boehringer sell a similar column packed with sephadex for probe purification
>their protocol uses swing out buckets rather than a fixed angle rotor.
>
>Excerpt from product spec sheet
>
>" In a fixed angle rotor, the sample is likely to slide down the sides of the tube instead of
>flowing through the medium. This results in poor retention of nucleotides and decreased recovery of
>sample "
>
>Just a thought
>
>Allison


I wish that was the problem, but I was using a swing out bucket rotor in the
experiment I described.  My guess is that the BioRad biospin frits are trapping
the plasmid sized DNA, but I don't have any proof for this interpretation
beyond what I mentioned already.

Any other experiences using BioRad biospin columns with kilobase sized DNA?


Song Tan
Institute for Molecular Biology and Biophysics
ETH-Honggerberg (Swiss Federal Institute of Technology)
8093 Zurich, Switzerland
email:  tan at aeolus.vmsmail.ethz.ch




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