By hcl7 on Thursday, December 5, 2002 - 08:57 am:
I’m rather a molecular biologist than an analyst but our team is using the Wave system (Denaturing HPLC for DNA analysis) from Transgenomic since 1998. We upgraded our system to the 3500 HighThroughput 6 months ago. This 3500HT system is expected to reduce elution times from 6 to 2 min without any quality loss. This is indeed the case for approx. 80% of our samples. Unfortunately, we also experience serious difficulties with some DNA amplicons that were well resolved with the previous system:
1) Wave-like (!) chromatograms (ie succession of small, broad peaks)
2) Lack of reproducibility (profile's variations for the same mutation over the series) and repetability (profile's variations for the same sample injected several times).
Although Transgenomic completely acknowledged these problems, they don't give us a piece of solution...
Please, share your experience if you encountered these troubles. Thanks!
By Anonymous on Thursday, December 5, 2002 - 02:47 pm:
No experience with your problem, but this sounds like pump problems / gradient mixing problems. What does Transgenomic say?
By hcl7 on Friday, December 6, 2002 - 05:19 am:
they didn't have any explanation for the moment, they just acknowledged the problems, telling us they will try to find out a solution...
Another Wave user advised me to increase re-equilibration time as it is extremly (too?) reduced with the HighThroughput version.
What's your opinion on this?
By hcl7 on Wednesday, January 15, 2003 - 06:40 am:
Thanks Anonymous,
you were right, it was a pump problem: if you experienced wavy chromatograms on a 3500 HT, check your pumps. They should be set to the “fast”position and not to the “slow” one…
By Anonymous on Friday, January 17, 2003 - 04:29 pm:
Thanks for getting back to us!!!
We sometimes give advice without ever getting feedback if it worked or if our guesses were correct
By Whompy Whopmperson on Thursday, June 5, 2003 - 12:15 pm: