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Qiaex problems (was Re: Gel Purification of DNA with Qiaex beads)

John Dixon jpcd0 at mole.bio.cam.ac.uk
Fri Jul 29 10:21:08 EST 1994


In article <318ac6$cfp at gazette.bcm.tmc.edu>, sc690763 at mbcr.bcm.tmc.edu
(Steven Chua) wrote:

> I am in the process of purifying a DNA fragment for injection into mice to
> generate transgenic mice and I am wondering what other methods are available
> to get a clean fragment with good yield that are suitable for injection... 
> 
> 
> With the qiaex beads that I am using, the yield I obtain is pathetic...my
> recovery is usually only about 10% of the original starting material... I have
> even called the company up for tips but my yield is still pretty low... When I
> used about 60ug of insert(the fragment I will be injecting) as my starting
> material, I only got back less than 6ug....
> 

I used Qiaex happily for 6 months during which I regularly got 90%+
recovery of 200bp-8kb fragments. Suddenly the Qiaex stopped working and I
only recovered feeble amounts. I have tried all sorts of potential
solutions without success and have now given up on Qiaex completely. I
would be delighted if somebody could explain what might have happened, we
have tried new kits without success either. 

Has anyone else had/overcome this problem?

-- 
John Dixon                     Lab 44 (223) 334131
Wellcome/CRC Institute         Fax 44 (223) 334134
Dept Genetics
Cambridge University           e-mail jpcd0 at mole.bio.cam.ac
United Kingdom



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