کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
7613774 1493597 2014 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Orthogonal ion pairing reversed phase liquid chromatography purification of oligonucleotides with bulky fluorophores
ترجمه فارسی عنوان
کروماتوگرافی مایع فازی متناوب جفت ارگوگنال، جبران سازی الیگونوکلئوتید با فلوئوروفورهای بزرگ
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی
A dual labeled oligonucleotide used as TaqMan® or 5′ nuclease probe for in vitro diagnostic has been purified through orthogonal ion-pairing reversed phase chromatography, using polymeric semi-preparative and preparative PRP-1 column. We studied the mechanism of separation of oligonucleotides using ion-pairing reversed phase chromatography. We found that elution profiles of dye labeled oligonucleotides can be controlled by use of specific ion-pairing reagents. Here, we report a method for purification of an oligonucleotide containing an internally positioned rhodamine dye using two orthogonal chromatographic steps, in which the primary step resolves mostly by differences in hydrophobicity by using a weak ion-pairing reagent, and a secondary step uses a strong ion-pairing reagent for separation of length variants. Purification is demonstrated for both 1 and 15 μmol scale syntheses, and amenable to further scale up for commercial lot production.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1329, 14 February 2014, Pages 78-82
نویسندگان
, , ,