Plasmid elution from filter problem
(by dprieto from fcien.edu.uy)
Wed Sep 17 14:40:27 EST 2008
Great info! Didn't know the type of paper was that critical. I will try
the high salt elution. Thank you!
Aawara Chowdhury wrote:
> 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 .....
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