High-performance capillary electrophoretic separation of double-stranded oligonucleotides using a poly-(ethylpyrrolidine methacrylate-co-methylmethacrylate)-coated capillary

Kerrilee Allan, Claire Lenehan, Dmitriy Khodakov, Hilton Kobus, Amanda Ellis

    Research output: Contribution to journalArticlepeer-review

    13 Citations (Scopus)

    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.

    Original languageEnglish
    Pages (from-to)1205-1214
    Number of pages10
    JournalElectrophoresis
    Volume33
    Issue number7
    DOIs
    Publication statusPublished - Apr 2012

    Keywords

    • Capillary electrophoresis
    • DNA
    • Oligonucleotides
    • Physically adsorbed coatings
    • Poly(ethylpyrrolidine methacrylate-co-methyl methacrylate) copolymer

    Fingerprint Dive into the research topics of 'High-performance capillary electrophoretic separation of double-stranded oligonucleotides using a poly-(ethylpyrrolidine methacrylate-co-methylmethacrylate)-coated capillary'. Together they form a unique fingerprint.

    Cite this