title: High-performance capillary electrophoretic separation of double-stranded oligonucleotides using a poly-(ethylpyrrolidine methacrylate-co-methylmethacrylate)-coated capillary.
abstract: Here we describe a capillary electrophoretic method for the separation of double-stranded oligonucleotides (ds-ODNs) ranging from 16-20 bp with 2 bp resolution using a low concentration of poly(ethylpyrrolidine methacrylate-co-methyl methacrylate) (PEPyM-co-PMMA) copolymer physically adsorbed to a capillary surface. Contrary to traditional DNA separations, we show that the ds-ODN with the highest molecular size eluted first and propose that this phenomena is due to a screening effect by the PEPyM-co-PMMA coating on the smaller ds-ODNs negative charge during elution. Key to the performance of this separation was a sample preparation time of less than 1 h and analysis time of 40 min. Repeatability of intraday migration time for the mixtures was typically < 1% relative standard deviation (n = 3). In addition, we demonstrate that the coating has an acceptable capillary lifetime of over 70 injections.
authors.affiliation: Flinders Centre for NanoScale Science and Technology, Flinders University, Adelaide, SA, Australia.
authors.forename: Kerrilee E
authors.initials: KE
authors.lastname: Allan
authors.forename: Claire E
authors.initials: CE
authors.lastname: Lenehan
authors.forename: Dmitriy A
authors.initials: DA
authors.lastname: Khodakov
authors.forename: Hilton J
authors.initials: HJ
authors.lastname: Kobus
authors.forename: Amanda V
authors.initials: AV
authors.lastname: Ellis
mesh_terms: D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry / Q000302:isolation & purification*; D011109:Polymethacrylic Acids / Q000737:chemistry*; D011758:Pyrroles / Q000737:chemistry*; D015203:Reproducibility of Results; D013696:Temperature
chemical_list: D009841:Oligonucleotides; D011109:Polymethacrylic Acids; D011758:Pyrroles