Regarding usage of waters Xbridge column with

[harish]

Hallo,
I have a akta 900 fplc and i want to seperate single strand oligos PCR PRODUCTS 20-100 bases with a waters xbridge 4.6*50 mm column. Can anyone say if these columns work at slow flowrates, company says ok..but i am not sure about modifications to be made and also i am getting a oven bulit and i dont know if can use a oven suitable for longer columns for this small one. My column is made of heating element and ventilator for cooling. can i just use a 2.5*150 mm column oven for my purpose..
sorry if it a long question,
i am not able to get over this problem for a long time to seperate ssDNA short fragments..
Thanks in advance,
Greetings,
Harish.

[Chris0000]

Hi

basically, the column will work, but RP columns tend to make a high backpressure while the FPLC systems normally are not able to tho work at high pressure.
You will have to slow down the flow in order to reduce pressure...so its not optimal but a possible way.

I would try to avoid to small particle sizes because of the backpressure and would not go smaller than 4.6 mm I.D. ( this is just a feeling, I dont know how precise the Äkta pump behaves at slow flowrates)

...and you might have to change some fittings in order to mount the column.

Best regards
Chris

[harish]

thanks for the advice. it works at low flowrates, but i cant seperate 2 samples in a single run. i have to do two runs. Can you suggest any possible gradient for this. buffers 20/80 ACN, 0.1M TEAA. i always see a peak just after injection, im not sure if it is a sample (it should be) or bubble.
Thanks for the help,
Greetings,
Harish.

[Uwe Neue]

Here is a poster with plenty of suitable information on the type of gradient that you need to run:
http://www.waters.com/webassets/cms/lib ... 2358en.pdf

If you can't get to it directly, go the Waters website, search in the corner for oligonucleotides, then select the information that you want (for example the first poster) and follow instructions.