Electrophoresis, Vol.26, No.23, 4449-4455, 2005
Fast separation of single-stranded oligonucleotides by capillary electrophoresis using OliGreen (R) as fluorescence inducing agent
The fast separation of oligonucleotide (oligos) sizing marker by CE using OliGreen (R) 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 x 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 x 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.
Keywords:capillary electrophoresis;ethidium bromide;high resolution;oligonucleotides;OliGreen (R);pluronic F127