Plasmid elution from filter problem

Aawara Chowdhury via methods%40net.bio.net (by aawara from pontiff-playground.org)
Wed Sep 17 08:17:29 EST 2008


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 .....

AC
-- 
Email: echo 36434455860060025978157675027927670979097959886449930P | dc


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