I'm looking for a system of DNA fragments analysis sold by Transgenomic co. This system allows to detect single base pair mutations in DNA fragment based on the separation of single-standed DNA (ssDNA) on a C18 RP column based on non-porous poly (styrene-divynylbenzene) particles at denaturing temperature (more than 55 celcius) using TEAA as ion pairing reagent. ssDNA is eluted in a linear gradient of acetonitrile.
My problem is the price of this apparatus, 105 000$ (US). What is the trick in this system to be so expensive? According to me it is a standard HPLC system with a UV detector. Is anybody see a problem if i use another HPLC system to perfom my analysis?
Moreover, gradient ion-pairing are not recommended (slow equilibration creating unstable system) Is anyboby know why this separation use acetonitrile gradient?
Tanks for Help.

Dear Mario,
I have no experience in the separation of DNA fragments, but we use regularly non-porous stationary phases for the separation of polypeptides. Using gradient methods NO special equipment is necessary! (Chromatographia 42(1996)595). For the separation of many - non polymeric - analytes we tried gradient elution AND ion pairing. We didn't found much difficulties (Chromatographia 50(1999)75).
Best regards.

While I am not directly involved in this area of chromatography, I have kept an eye on participants in this commercial arena. I believe that Varian sells a product which is competitive. Also, at the most recent WCBP meeting in San Francisco I saw some marketing literature from Agilent indicating that they had developed a silica based version of this column. If I were you, I would contact each of these companies to determine what sort of products they are offering and their level of expertise in supporting their product.

I was one of the first people to buy the Transgenomic system in the US. The system is robust and very good for mutation detection. The columns are exceptional. However the price is high. Our justification for purchasing the system was "that is the price you need to pay to be a leader in the field". We analyse in excess of 12 genes, some up to 25 exons in over 500 patients (thats a lot of samples)

After having systems now for over two years, we would not look back. The HPLC is manufactured by Hitachi Instruments. We have worked this machine so hard and have not encountred any problems. In fact, I changed the pump seals at the end of last year after 17000 injections. We replaced the lamp a couple of times (remember we operate it 24 hrs a day, 7 days a week).

As a recommendation, buy the Transgenomic system. I did try all the alternatives, but nothing was as good as this system. I would also suggest swapping out the 500uL syringe for a 100uL and purchasing a buffer wash kit (both available from Hitachi Instruments). Your system will perform much better !!!

Hope this helps you.

I would like to ask the same question as Mario again- why should I buy a completely new expensive system for a new method? Is there any special reason for it? What about to buy just the column (from Transgenomics or the Zorbax Eclipse dsDNA) and connect it to "standard" HPLC system.Have anybody tried the DHPLC method on such "standard" system with the Transgenomics or Zorbax or another columns (some authors reported the comparisson of Transgenomic and Micra NPS silica column)? Why should be the system inert?
Thanks for suggestions

The HP Eclipse is not as robust as the Transgenomic DNA Sep for this analysis (unless they've changed it since we ran our trials). Base pair separation is not as good !!! (I believe the DNASep column has at least 2 bp resolution, but you might want to check with them. Transgenomic also provide excellent support for their applications.

HAllo!
Can anyone recommend a column for analysing fermentation broth by HPLC. Especially organic acids and carbohydrats are of interest, detection by RI-detector. At the moment I am using an ORH801 column (now with Transgenomics label I think) but I do not know if this is really the best for this purpose (and of course not the cheapest.) My dealer offered me as alternatives a benson polymeric organic acid column, ICSept ICE-ION-300, TG-Coregel 87H Ionexclusion H+form, or CARBOSep H plus.
Comments please!
Many thanks in advance.
Gerd