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Analysis and Purification of Synthetic Oligonucleotides by Reversed-Phase High-Performance Liquid Chromatography with Photodiode Array and Mass Spectrometry Detection

✍ Scribed by Martin Gilar

Publisher
Elsevier Science
Year
2001
Tongue
English
Weight
113 KB
Volume
298
Category
Article
ISSN
0003-2697

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✦ Synopsis

Native and modified synthetic oligonucleotides were purified by reversed-phase HPLC using volatile ionpairing mobile phases. Purification of 10 -90 nmol of oligonucleotides in a single injection was demonstrated using a 4.6 Ψ‹ 75-mm HPLC column packed with porous 2.5 m 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.

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