Purification of dye-labeled oligonucleotides by ion-pair reversed-phase high-performance liquid chromatography.
J Chromatogr B Analyt Technol Biomed Life Sci
; 783(1): 61-72, 2003 Jan 05.
Article
em En
| MEDLINE
| ID: mdl-12450525
ABSTRACT
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 micrometer 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 (Cy3) dyes, as well as dually-labeled TaqMan probes. Purification of a 0.1-micromole oligonucleotide synthesis in a single injection was demonstrated.
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Base de dados:
MEDLINE
Assunto principal:
Oligonucleotídeos
/
Cromatografia Líquida de Alta Pressão
/
Corantes Fluorescentes
Idioma:
En
Ano de publicação:
2003
Tipo de documento:
Article