HPLC Purification

The DNA Synthesis Facility at the Biotech Center now has the ability to perform HPLC purifications upon user request. We utilize a reverse phase column that elutes DNA by both increasing length and hydrophobicity. In contrast to cartridge purification, this technique bypasses the need for the trityl group and the harsh reagents required to remove the trityl post-purification. We can separate the full length oligo from the N-1 peak up to a length of about 40 nucleotides using our standard solvent system. For longer oligos other solvent systems can be employed (such a request comes with a significant cost increase), though the system as presently configured still generates 95+% purity in longer oligos. Standard HPLC purification for such oligos will be $60/sample, regardless of whether purification is requested on 40nmole, 0.2 µmole or a1.0 µmole scale.

Singly labeled oligos (e.g., 5' FAM, HEX, TET, etc.) also utilize standard solvent systems since the label is hydrophobic and allows separation from unlabeled failure sequences. Cost for purification for these will also be $60/oligo. Multiply labeled oligos (e.g., those used in TAQMAN assays or other FRET applications, or labeled with multiple spacers, etc.) require more tweaking and analytical runs prior to the preparative run to insure collection of the correct final product. Pricing for these is subsequently higher, $85/oligo. As above, pricing is independent of scale.

In addition to the purified product, user will be provided with documentation including a digitally generated run log, and an analog run profile showing the collected fractions. If applicable, a UV/VIS spectrophotometer scan will be provided to show incorporation of the correct fluorescent labels into the purified product.

As always we can consult with users on any preferences they have. Contact us by phone or email and we'll do our best to help or answer any questions.

To request HPLC purification simply click on the HPLC button from the list of purification options provided in the Online Order Form.