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AssigneeApplera Corporation
InventorsRamakrishna Madabhushi, Steven Menchen, J. Efcavitch, Paul Grossman
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The invention provides uncharged water-soluble silica-adsorbing polymers for suppressing electroendoosmotic flow and to reduce analyte-wall interactions in capillary electrophoresis. In one aspect of the invention, one or more of such polymers are employed as components of a separation medium for the separation of biomolecules, such as polynucleotides, polysaccharides, proteins, and the like, by capillary electrophoresis. Generally, such polymers are characterized by (i) water solubility over the temperature range between about 20° C. to about 50° C., (ii) concentration in a separation medium in the range between about 0.001% to about 10% (weight/volume), (iii) molecular weight in the range of about 5×103 to about 1×106 daltons, and (iv) absence of charged groups in an aqueous medium having pH in the range of about 6 to about 9. In one embodiment, polymers of the invention are selected from the group consisting of polylactams, such as polyvinylpyrrolidone; N,N-disubstituted polyacrylamides; and N-substituted polyacrylamides. In accordance with the method of the invention, a sufficient amount of polymer adsorbs to the capillary surface to establish a zone of high viscosity that shields the analyte from the wall and impedes the movement of an electrical double layer under an electric field.
A method of suppressing electroendoosmotic flow in capillary electrophoresis, the method comprising providing a separation medium containing one or more uncharged water-soluble silica-adsorbing polymers having (i) water solubility in a temperature range between about 20° C. and about 50° C., (ii) a concentration in the separation medium in a ranae between about 0.001% and about 10% (weight/volume), (iii) a molecular weight in the range between about 5×103 and about 1×106 daltons, and (iv) an absence of charged groups in an aqueous medium having a pH in the range between about 6 and about 9.
A method of suppressing wall-analyte interaction in capillary electrophoresis, the method comprising providing a separation medium containing one or more uncharged water-soluble silica-adsorbing polymers having (i) water solubility in a temperature range between about 20° C. and about 50° C., (ii) a concentration in the separation medium in a range between about 0.001% and about 10% (weight/volume), (iii) a molecular weight in the range between about 5×103 and about 1×106 daltons, and (iv) an absence of charged groups in an aqueous medium having a pH in the range between about 6 and about 9.
A composition for separating polynucleotides by capillary electrophoresis, the composition comprising:
A method of separating different-sized polynucleotides by electrophoresis in an uncoated silica capillary, the method comprising the steps of:
Dependencies are inferred from the claim text.