By Bill on Monday, May 24, 2004 - 10:12 pm:
I want to separate the four nucleotides A,T,C,G by HPLC, to determine the base composition of a bacterial genome. I am new to HPLC and would like to ask what kinds of column to use?
By Patrik Appelblad on Tuesday, May 25, 2004 - 01:03 pm:
Dear Bill,
An alternative is to use our ZIC-HILIC phase.
Below you'll find an hyperlink to an application note, in the last LCGC Europe magazine, dealing with this particular topic.
http://www.lcgceurope.com/lcgceurope/data/articlestandard/lcgceurope/192004/94777/article.pdf
By X.L. on Tuesday, May 25, 2004 - 06:04 pm:
You also can try polar-embedded RP columns, such as Symmetry Shiled C18 from Waters, Agilent Bonus RP from Agilent, or Acclaim PolarAdvantage from Dionex. I have experience with all three and am pretty sure you can baseline separate 5 nucleotides. The mobile phases I used are 1)2.5mM H3PO4, or 2) 20 mM phosphate buffer, pH3.2. UV at 254 nm.
Email me if you need more information.
Good luck!
By Zelechonok on Tuesday, May 25, 2004 - 08:26 pm:
Here is another way of separating 4 nucleotides and uracil using mixed mode column
http://allsep.com/makeChr.php?chr=Chr_013