Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 783, Issue 1, Pages 61-72Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S1570-0232(02)00490-7
Keywords
genotyping; oligonucleotides
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Singly- and dually-labeled synthetic oligonucleotides were purified by ion-pair reversed-phase high-performance liquid chromatography using a 50x4.6-mm column packed with porous, 2.5 mum C-18 sorbent. We studied the mechanism of dye-labeled oligonucleotide retention in order to improve the quality of purification. By-products of oligonucleotide synthesis were characterized by liquid chromatography with mass spectrometry detection (LC-MS). We purified oligonucleotides labeled with 6-carboxyfluorescein (6FAM), hexachlorofluorescein (HEX), tetrachlorofluorescein (TET), carboxytetramethylrhodamine (TAMRA) and indodicarboxycyanine (Cy(TM)3) dyes, as well as dually-labeled TaqMan(TM) probes. Purification of a 0.1-mumole oligonucleotide synthesis in a single injection was demonstrated. (C) 2002 Elsevier Science B.V. All rights reserved.
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