Qiagen Quiaquick : reliable? pure enough for Autoseq?

Lely Chow lelyc at u.washington.edu
Thu Sep 5 20:06:00 EST 1996


Hi,

I don't know if this will help, but we just started using the Qiaquik kit
for gel purification, and the problem seems to be that there is EtOH
contamination in the eluate.  To solve the problem, do the spin after the
last wash (contains EtOH) for much longer than they say, or better yet,
speedvac the sample on heat to get rid of the EtOH.  We have called the
company, and they are aware of this problem.  This was their suggestion,
and seems to work just fine.

--Lely




On 23 Aug 1996, Matt Hudson wrote:

> Does anyone there notice variability in the purity and yield from the
> Quiaquick kits ('specially the gel kit). Sometimes I get nice DNA with
> 80% yield and a lovely spec. with good 260:280. More often, and
> especially recently, since the tops of the little purple columns went
> flat instead of dimply, the yield is dismal and there is a funny little
> shoulder on the spec. at 220-230nm. I follow their protocol exactly each
> time, including the optional bits for purer DNA.
>
> My problems (read only if genuinely interested)
> I am trying to sequence a genomic coding region where I reckon there is
> a single-base mutation. I've got lots of primers producing overlapping
> products from it which I am sequencing directly, using PE dye
> terminators and primers (there are PE primer sequenses at the 5' end of
> the amplimers which makes them BIG - 45bp). The trouble is not all the
> primers work that well, so I can't just remove primers and nucleotides
> and add the pcr reaction - I need to gel purify the band. Getting DNA of
> decent enough quality to do this from a gel has eluded me for some time
> - I do get sequence, but it's dodgy with short reads and highly
> inaccurate. Not what you need for spotting one different base {:-(}.
>
> Thank you anyone out there who takes pity on me
> Yours worried
>
> Matt Hudson
> Botany Dept
> Leicester University
> Leicester LE1 7RH
> UK
>
> tel. (UK) 0116 252 3399
> meh2 at le.ac.uk
>
>
>
>




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