Premium
Fast separation of single‐stranded oligonucleotides by capillary electrophoresis using OliGreen® as fluorescence inducing agent
Author(s) -
Zhang Jun,
Liang Dehai,
He Weidong,
Wan Fen,
Ying Qicong,
Chu Benjamin
Publication year - 2005
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200500099
Subject(s) - chromatography , capillary electrophoresis , ethidium bromide , oligonucleotide , chemistry , resolution (logic) , analytical chemistry (journal) , electrophoresis , urea , fluorescence , capillary action , materials science , dna , biochemistry , optics , physics , artificial intelligence , computer science , composite material
The fast separation of oligonucleotide (oligos) sizing marker by CE using OliGreen® and including effects due to the concentration of separation medium and urea denaturant is presented. OliGreen dye is found to be more sensitive than ethidium bromide (by a factor of about 6 based on S/N considerations) for the oligos' separations. Higher concentration of F127 in 1×Tris‐boricacid‐EDTA (TBE) up to 30% w/v leads to better resolution of oligos separations. The addition of urea into the separation medium decreases the sensitivity. With an optimized running condition, the oligos sizing marker could be successfully separated with 1‐base resolution within 1.3 min by using 30% w/v F127/1×TBE solution as the separation medium at an applied electric field of 800 V/cm in a 3 cm long capillary, the fastest capillary gel electrophoresis separation with high resolution reported to date for oligos in the similar size range.