Journal
ANALYTICAL BIOCHEMISTRY
Volume 298, Issue 2, Pages 196-206Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1006/abio.2001.5386
Keywords
oligonucleotide purification; ion-pair reversed-phase; HPLC; mass spectrometry; fluorescent primers; genotyping
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Native and modified synthetic oligonucleotides were purified by reversed-phase HPLC using volatile ion-pairing mobile phases. Purification of 10-90 nmol of oligonucleotides in a single injection was demonstrated using a 4.6 x 75-mm HPLC column packed with porous 2.5 mum C18 sorbent. Separation of target products from N-1 failure fragments was achieved for oligonucleotides in the 4- to 60-mer size range. We employed a combination of absorbance and mass spectrometry detection to identify by-products of oligonucleotide synthesis. This method was also employed for analysis and purification of fluorescently labeled oligonucleotides. (C) 2001 Academic Press.
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