Jennifer M. Clyne

Bio: Jennifer M. Clyne is an academic researcher from Chiron Corporation. The author has contributed to research in topics: Oligonucleotide & Nucleic acid. The author has an hindex of 6, co-authored 13 publications receiving 518 citations. Previous affiliations of Jennifer M. Clyne include Siemens.

A comparison of non-radioisotopic hybridization assay methods using fluorescent, chemiluminescent and enzyme labeled synthetic oligodeoxyribonucleotide probes.

Abstract: N4-[N-(6-trifluoroacetylamidocaproyl)-2-aminoethyl]-5'-O-dimethoxy trityl -5-methyl-2'-deoxycytidine-3'-N,N-diisopropyl-methylphosphoramidite++ + has been synthesized. This N4-alkylamino deoxycytidine derivative has been incorporated into oligonucleotide probes during chemical DNA synthesis. Subsequent to deprotection and purification, fluorescent (fluorescein, Texas Red and rhodamine), chemiluminescent (isoluminol), and enzyme (horseradish peroxidase, alkaline phosphatase) labels have been specifically incorporated. Detection limits of the labels and labeled probes were assessed. Also, the detection limits and nonspecific binding of the labeled probes in sandwich hybridization assays were determined. The enzyme modified oligonucleotides were found to be significantly better labeling materials than the fluorescent or chemiluminescent derivatives, providing sensitivities comparable to 32P-labeled probes.

Improved amplified nucleic acid hybridization assays for hbv.

Abstract: Amplified solution-phase sandwich nucleic acid hybridization assays for HBV in which (1) analyte is hybridized in solution with sets of amplifier probe oligonucleotides and capture probe oligonucleotides each having a first segment that is complementary to a consensus HBV ds region sequence based on a multiplicity of HBV subtypes and a second segment that is complementary to a unit of an oligonucleotide multimer and an oligonucleotide bound to a solid phase, respectively (2) the resulting product is reacted with the oligonucleotide bound to a solid phase (3), the resulting product is washed to remove unbound materials (4), the bound materials are reacted with the multimer (5), the bound materials are washed to remove unbound multimer (6), the bound materials are reacted with a labeled probe complementary to the oligonucleotide units of the multimer.

Nucleic acid hybridization assay for hepatitis B virus DNA

Abstract: Linear or branched oligonucleotide multimers useful as amplifiers in biochemical assays which comprise (1) at least one first single-stranded oligonucleotide unit that is complementary to a single-stranded oligonucleotide sequence of interest, and (2) a multiplicity of second single-stranded oligonucleotide units that are complementary to a single-stranded labeled oligonucleotide. Amplified sandwich nucleic acid hybridizations and immunoassays using the multimers are exemplified.

Bifunctional blocked phosphoramidites useful in making nucleic acid mutimers

Abstract: Linear or branched oligonucleotide multimers useful as amplifiers in biochemical assays which comprise (1) at least one first single-stranded oligonucleotide unit that is complementary to a single-stranded oligonucleotide sequence of interest, and (2) a multiplicity of second single-stranded oligonucleotide units that are complementary to a single-stranded labeled oligonucleotide. Amplified sandwich nucleic acid hybridizations and immunoassays using the multimers are exemplified.

Selective Colorimetric Detection of Polynucleotides Based on the Distance-Dependent Optical Properties of Gold Nanoparticles

Abstract: A highly selective, colorimetric polynucleotide detection method based on mercaptoalkyloligonucleotide-modified gold nanoparticle probes is reported. Introduction of a single-stranded target oligonucleotide (30 bases) into a solution containing the appropriate probes resulted in the formation of a polymeric network of nanoparticles with a concomitant red-to-pinkish/purple color change. Hybridization was facilitated by freezing and thawing of the solutions, and the denaturation of these hybrid materials showed transition temperatures over a narrow range that allowed differentiation of a variety of imperfect targets. Transfer of the hybridization mixture to a reverse-phase silica plate resulted in a blue color upon drying that could be detected visually. The unoptimized system can detect about 10 femtomoles of an oligonucleotide.

Very large scale immobilized polymer synthesis

Abstract: A method and apparatus for preparation of a substrate containing a plurality of sequences. Photoremovable groups are attached to a surface of a substrate. Selected regions of the substrate are exposed to light so as to activate the selected areas. A monomer, also containing a photoremovable group, is provided to the substrate to bind at the selected areas. The process is repeated using a variety of monomers such as amino acids until sequences of a desired length are obtained. Detection methods and apparatus are also disclosed.

Method of detecting nucleic acids

Abstract: The present invention provides method and apparatus for sequencing, fingerprinting and mapping biological macromolecules, typically biological polymers. The methods make use of a plurality of sequence specific recognition reagents which can also be used for classification of biological samples, and to characterize their sources.

Combinatorial strategies for polymer synthesis

Abstract: A method and device for forming large arrays of polymers on a substrate ( 401 ). According to a preferred aspect of the invention, the substrate is contacted by a channel block ( 407 ) having channels ( 409 ) therein. Selected reagents are delivered through the channels, the substrate is rotated by a rotating stage ( 403 ), and the process is repeated to form arrays of polymers on the substrate. The method may be combined with light-directed methodolgies.

Fluorescence resonance energy transfer and nucleic acids.

Abstract: Publisher Summary Fluorescence resonance energy transfer (FRET) is a spectroscopic process by which energy is passed nonradiatively among molecules over long distances. The “donor” molecule, which must be a fluorophore, absorbs a photon and transfer this energy nonradiatively to the “acceptor” molecule. Energy can be transferred over distances on the order of common macromolecular dimensions. This chapter discusses the applications of FRET to nucleic acid molecules conjugated to dye molecules; but in general, the donor-acceptor pairs can be free in solution, one or both bound to a macromolecule, or be an inherent part of the structure. The chapter discusses FRET specifying the physical parameters that are available from a variety of fluorescence measurements. In the chapter, some basic considerations and experimental procedures are described that are necessary to make reliable FRET measurements. It also discusses some difficulties that can arise in the experimental proceedings and data analysis and the sources of experimental error are pointed out.