Plasmid elution from filter problem

[Pow Joshi]

2008/9/17 Aawara Chowdhury <aawara from pontiff-playground.org>

> In <mailman.1501.1221496835.3533.methods from net.bio.net>,
>  Daniel Prieto <dprieto from fcien.edu.uy> wrote:
>
> > Hi all, I am trying to elute some plasmids from a whatman paper but do
> > not seem to achieve it. I tried the classic 50 uL water, 5 minutes at RT
> > and spinned down the paper. I am usually quite successful using this
> > technique with other plasmids. I "eluted" them this way and tried to
> > transform (2 different methods) with no results, I ran the DNA on an
> > agarose gel but could not see any band. The guys who sent it to me told
> > me they were concentrated enough to see them quite well with EtBr. I
> > have had them for almost a year in the filter prior to my attempts to
> > elute them, but I don't see much of a problem in that. Can someone give
> > me some suggestions to improve my yield with the elution?
>
> Are you sure they're on Whatman filter paper, and not on Whatman DE-81
> paper?  I was recently sent some plasmids spotted on what was assumed
> to be filter paper - I could not elute them with water or TE, but they
> came off in high salt (1M sodium acetate, pH 5.5), and I transformed
> them after drop-dialysis.
>
> About a week after this happened, I received an email from the colleague
> I had requested the plasmids from indicating that his new technician
> had been sending plasmids spotted on DE81 paper and not 3mm paper, and
> that they would be sending the plasmids again .....


Thanks AC, this is good to know. One of my colleagues had similar trouble an
year ago, and I am now wondering if we should've tried the high salt/pH way.


Pow


>
>
> AC
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