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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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Past Experience
Posted by: EANON (IP Hidden, Junior member, 19)
Date: July 8, 2005 08:24PM
I recently spent over a week troubleshooting a Maxiprep for DNA purification from E. Coli cells- I was getting an extremely low yield of DNA. To make a long story short, I changed the elution buffer, temperature and volume of elution buffer, volume of bacterial grow-up, lysis time, I used someone else's kit, I had someone watch me perform the protocol, and tried several other tactics.
I finally found that the problem was with the 70% ethanol I used in the wash. It was either mislabeled or contaminated, in any case, my maxipreps are now acceptable. I am very interested in hearing what problems others have had with similar simple protocols - maxipreps, co-IPs, Westerns, etc - and what the problem turned out to be.
Re: Past Experience
Posted by: Larman (IP Hidden, Unregistered user, )
Date: July 9, 2005 12:35PM
It is always a good idea to make all buffers fresh if something goes terribly wrong in a biochemistry experiement. It is the most efficient way of troubleshooting.
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