Prolinx Incorporated

Karin A Hughes

Mark L Stolowitz

Edward A Kesicki

Kevin P Lund

Robert J Kaiser

Douglas A Spicer

James E Mahoney

Amy L Springer

Carl H D Weissman

This invention provides methods for removing unincorporated dye-labeled molecules from a mixture that includes dye-labeled polynucleotides or other polymers and the unincorporated dye-labeled molecules. The methods involve adsorbing the unincorporated dye-labeled molecules into a plurality of particles that are made up of one or more porous hydrophobic materials that are encapsulated in a hydrophilic matrix.

Removal of dye-labeled dideoxy terminators from DNA sequencing reactions

Phenyldiboronic acid reagents and complexes

This invention provides methods for removing unincorporated fluorescent dye-labeled molecules from a mixture that includes fluorescently-labeled polynucleotides and the unincorporated fluorescent dye-labeled molecules. The methods involve adsorbing the unincorporated fluorescent dye-labeled molecules into a plurality of particles that are made up of one or more porous hydrophobic materials that are encapsulated in a hydrophilic matrix.

Reagents suitable for the modification of a bioactive species for the purpose of incorporating one or more phenyldiboronic acid (PDBA) moieties for subsequent conjugation to a different (or the same) bioactive species having one or more pendant boronic compound complexing moieties of the general formula of General Formula I, ##STR1## wherein group R is a reactive electrophilic or nucleophilic moiety suitable for reaction of the PDBA reagent with a bioactive species. Group Z is a spacer selected from a saturated or unsaturated chain up to about 0 to 6 carbon equivalents in length, an unbranched saturated or unsaturated chain of from about 6 to 18 carbon equivalents in length with at least one intermediate amide or disulfide moiety, and a polyethylene glycol chain of from about 3 to 12 carbon equivalents in length. Group Q is a linkage that includes one of amide, ether and thioether moieties. Group R is preferably selected from, but not limited to, one of acrylamide, bromo, bromoacetamide, chloro, chloroacetamide, dithiopyridyl, hydrazide, N-hydroxysuccinimidyl ester, N-hydroxysulfo-succinimidyl ester, imidate ester, imidazolide, iodo, iodoacetamide, maleimide, amino and thiol moieties. Group Z is preferably an unbranched alkyl chain of the general formula (CH.sub.2)n, wherein n=1 to 6. Group Q is preferably selected from one of NHCO, CONH, NHCOCH.sub.2, CONHCH.sub.2, O, OCH.sub.2, S and SCH.sub.2 moieties.

Boron compound complexing reagents, boron compound complexes, and methods of synthesizing these reagents and complexes are disclosed. These reagents and complexes include those shown in General Formula CIII, General Formula CIV, and General Formula CVI. In one embodiment, the reagents of General Formula CIII may be used to produce, after condensation with a bioactive species (BAS), the reagent of General Formula CIV. The reagent of General Formula CIV may be used to form a complex with a boron compound, such as a complex shown in General Formula CVI. ##STR1##

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Bothell, WA, United States of America

Karin A Hughes