Abstract
SELEX (systematic evolution of ligands by exponential enrichment) is a process for isolating DNA or RNA sequences with high affinity and selectivity for molecular targets from random sequence libraries. These sequences are commonly referred to as aptamers. The process typically requires 10–15 cycles of enrichment, PCR amplification and nucleic acid purification to obtain high-affinity aptamers. We have demonstrated that using capillary electrophoresis (CE) as an enrichment step greatly improves the efficiency of the process. CE–SELEX is capable of isolating high-affinity aptamers in as little as 2–4 rounds of selection, shortening the process time from several weeks to as little as a few days.
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References
Joyce, G.F. (1989) Amplification, mutation, and selection of catalytic RNA. Gene 82, 83–87.
Ellington, A. D. and Szostak, J. W. (1990) In vitro selection of RNA molecules that bind specific ligands. Nature 346, 818–822.
Tuerk, C. and Gold, L. (1990) Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249, 505–510.
Famulok, M., Mayer G., and Blind, M. (2000) Nucleic acid aptamers-from selection in vitro to applications in vivo. Acc. Chem. Res. 33, 591–599.
Hamula, C.L.A., Guthrie, J.W., Zhang, H., Li, X.-F. and Le, X. C. (2006) Selection and analytical applications of aptamers. Trends. Analyt. Chem.. 25, 681–691.
Bunka, D.H.J. and Stockley, P.G. (2006) Aptamers come of age – at last. Natl. Rev. Microbiol.. 4, 588–96.
Tombelli, S., Minunni, M. and Mascini, M. (2005) Analytical applications of aptamers. Biosens. Bioelectron. 20, 2424–2434.
Mendonsa, S.D. and Bowser, M.T. (2004) In vitro evolution of functional DNA using capillary electrophoresis. J. Am. Chem. Soc. 126, 20–21.
Mendonsa, S.D. and Bowser, M.T. (2004) In vitro selection of high-affinity DNA ligands for human IgE using capillary electrophoresis. Anal. Chem. 76, 5387–5392.
Mosing, R.K., Mendonsa, S.D. and Bowser, M.T. (2005) Capillary electrophoresis-SELEX selection of aptamers with affinity for HIV-1 reverse transcriptase. Anal. Chem., 77, 6107–6112.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Mosing, R.K., Bowser, M.T. (2009). Isolating Aptamers Using Capillary Electrophoresis–SELEX (CE–SELEX). In: Mayer, G. (eds) Nucleic Acid and Peptide Aptamers. Methods in Molecular Biology™, vol 535. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-557-2_3
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DOI: https://doi.org/10.1007/978-1-59745-557-2_3
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Publisher Name: Humana Press, Totowa, NJ
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