Fast separation of single-stranded oligonucleotides by capillary electrophoresis using OliGreen as fluorescence inducing agent.
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 1xTris-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 F127/1xTBE 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.
Zhang
Jun
J
Department of Chemistry, Stony Brook University, Stony Brook, NY 11794-3400, USA.
Liang
Dehai
D
He
Weidong
W
Wan
Fen
F
Ying
Qicong
Q
Chu
Benjamin
B
D002021:Buffers; D004277:DNA, Single-Stranded / Q000737:chemistry; D019075:Electrophoresis, Capillary; D004996:Ethidium; D005456:Fluorescent Dyes / Q000737:chemistry*; D009841:Oligonucleotides / Q000302:isolation & purification*; D020442:Poloxamer; D012680:Sensitivity and Specificity; D014508:Urea
D002021:Buffers; D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D009841:Oligonucleotides; D020442:Poloxamer; D014508:Urea; D004996:Ethidium