Purification of DNA Oligonucleotides Using Oasis HLB 96-Well Extraction Plates

Library Number:
990854
Author(s):
Martin Gilar;Edouard S.P. Bouvier
Source:
ISPPP''99; Delray Beach FL; 1 Nov 1999
Content Type:
Posters
Content Subtype:
ISPPP
SPE Format:
SPE:
Oasis HLB 30 mg 96-well Plate Oasis HLB Prototype 9 micron 30 mg 96-well Plate
Sorbent:
Compounds:
DNA Oligonucleotides, tritylated
Matrix:
Synthesis mixture
Analytical Techniques:
Column:
XTerra MS 18 2.5 µm Steel 4.6 mm 50 mm XTerra MS 18 2.5 µm Steel 4.6 mm 75 mm Oasis® HLB 5 µm Prototype Column - steel 2.1 mm 20 mm SymmetryShield(™) RP8 5 µm Steel 3.9 mm 50 mm
Abstract: Purification of target oligodeoxyribonucleotides from DNA synthesis was developed based on a solid phase extraction ( trityl on purification) using a 96-well Oasis® HLB extraction plate. The Oasis® HLB sorbent combines excellent pH stability and a high loading capacity allowing for single step purification of 0.2 micromole scale synthesis. After washing failure sequences off, the oligonucleotide trityl group is cleaved in situ with 2% TFA, and target oligonucleotide is eluted with acetonitrile/0.36 mM triethylamine acetate, pH 11.3 (10/90, v/v). Typical yield of purified product is 75-95 %. Final purity, measured by capillary gel electrophoresis, was found to be 90 % or greater. Alternatively, >99 % purity oligonucleotides can be obtained using RP-HPLC Trityl off method with an XTerra™ MS C18 column.

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