Abstract:
The present invention relates to methods for the optical mapping and subsequent analysis, such as sequencing of polynucleotides, such as genomic DNA, in microchannel devices.
Abstract:
Methods of generating a nucleic acid signature for identifying particles associated in a partition are provided. In one aspect, the method comprises: partitioning a sample into a plurality of partitions comprising a particle comprising a solid support surface, the solid support surface having a plurality of oligonucleotide primers conjugated thereon, wherein the oligonucleotide primers comprise a barcode sequence, and wherein the partitions have 0, 1, or more than 1 particles per partition; providing in a partition a substrate comprising a barcode sequence or repeating clonal barcode sequences; and in the partition, associating a first particle conjugated to oligonucleotide primers comprising a first barcode sequence and a second particle conjugated to oligonucleotide primers comprising a second barcode sequence to a barcode sequence from the substrate, thereby generating a nucleic acid signature for the particles in the partition.
Abstract:
Photoblocked probes are disclosed including a specific hydrolysis probe having a first nucleic acid sequence complementary to a target having a second nucleic acid sequence, a first and a second interactive labels, a 5' end and a 3' end, and one or more photocleavable moieties coupled to one or more nucleotides of the specific hydrolysis probe, wherein the photocleaveable moiety interferes with the hybridization of the specific hydrolysis probe with the region of the amplification product. Also disclosed are PCR methods for the detection of the presence or absence of a target nucleic acid in a sample utilizing the photoblocked probes, as well as kits.
Abstract:
The invention relates to a method that utilizes nucleic acid deactivating reagents for the deactivation of contaminating nucleic acids, and thermophilic proteinases for the extraction of nucleic acids in a closed-system to be used in tandem with methods for the amplification of target nucleic acids present in a sample. The combined method enables simplified, temperature-controlled devices to be used for accurate, streamlined testing at the point of care for a wide variety of applications in the medical, industrial, environmental, quality control, security and research fields.
Abstract:
The invention relates to a novel method for obtaining DNA metasegment comprising ligating adjoining DNA fragments at least 10 Kb in size containing at least one overlapping region.
Abstract:
Photoreactive phosphoramidites useful for attaching photoreactive sites to nucleic acids and oligonucleotides are synthesized. The resultant nucleic acid or oligonucleotide probes incorporating the photoreactive sites are then attached to a polymer-coated support by a [2 + 2] cycloaddition to form a microarray.
Abstract:
A process is disclosed for generating at least one partially double-stranded target nucleic acid, which contains at least one single-stranded region at a tenninal end. The process comprises the steps of (a) providing at least one primer, P1, containing at least one labile nucleotide; (b) combining at least one target nucleic acid sequence with P1 to generate a double-stranded polynucleotide containing at least one labile nucleotide; (c) exposing the double-stranded polynucleotide to conditions that promote single-strand cleavage of the polynucleotide at the site of the at least one labile nucleotide of primer P1; and (d) exposing the cleaved polynucleotide to conditions that promote the dissociation of the cleaved portions of primer PI from a terminal end. The labile nucleotide may be dUTP, wherein the single-stranded cleavage of the polynucleotide at the site of the labile nucleotide occurs by treatment with uracil N-glycosylase.
Abstract:
A method detects binding of molecules, advantageously without tagging molecules in the sample. A sensor is used in which is included a single stranded nucleic acid sequence and a photoluminescent material in respective layers. After the sensor is exposed to a biological sample for sufficient time for its single stranded nucleic acid sequence to bind to a material of interest, photoluminescence from the sensor can be measured. An apparatus for tagging-free detection of binding of molecules also is provided. Methods of making tagging-free sensors are provided. Also, tagging-free methods to detect binding of antigens and related devices are disclosed.