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Poor resolution when isolating ssDNA?

Poor resolution when isolating ssDNA? - Protocols and Methods Forum

Poor resolution when isolating ssDNA? - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 06-12-2008, 09:56 PM
Evan Reed
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Default Poor resolution when isolating ssDNA?



Hej,

I am trying to isolate ssDNA after binding dsDNA oligos with
biotinylated antisense strands to strep microbeads, denaturing with
NaOH, then adding the supernatant containing the sense ssDNA to a
NAP-5 sephadex desalting column. Unfortunately, I am getting terrible
resolution when I try to elute on the column -- even after 2 mL of
elution, I am still getting detectable levels of ssDNA in the eluate
(dsDNA oligos of the same length elute just fine in 0.5 mL, with
acceptable resolution). The amount of DNA loaded into the column
should be well below the binding capacity so I am not sure why this is
happening. Has anyone done this before? Is there a better way to
purify the ssDNA instead of this column?

Thanks,

ER

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Old 06-17-2008, 06:12 AM
cheetah.zao@gmail.com
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Default Poor resolution when isolating ssDNA?

70% enthenol precipitation in the presence of 50 mM MgCl2 may work.



On Jun 13, 5:56 am, Evan Reed <[Only registered users see links. ]> wrote:

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