Abstract: Image processing for certain sequencing technologies requires data processing algorithms that provide fast sequence detection with low error rates. Methods and apparatus for performing image analysis for identifying nucleotide incorporations includes performing an image segmentation procedure on a plurality of data sets to identify sample objects and to create segmented data sets for each of the data sets. Each data set represents a sample image that includes a plurality of pixel locations and intensity data associated with each of the pixel locations. The segmented data sets represent identified sample objects for each one of the sample image data sets. An image registration procedure is performed on the segmented data sets to align the identified sample objects and to create data representative of the aligned identified sample objects. A strand formation procedure is then performed on the data representative of the aligned identified sample objects to identify nucleotide incorporations.

Abstract: Methods of the invention comprise methods and devices for nucleic acid sequence determination. Generally, the invention relates to preparing a substrate for sequencing a target nucleic acid.

Abstract: Single molecule technologies generally require sensitive optical detection and the ability to operate at multiple temperatures simultaneously in different parts of the instrument. The system for controlling the temperature of a microfluidic device and methods for controlling the temperature of sequencing reactions includes a chamber for receiving a microfluidic device, a heating control device in fluid communication with the chamber for delivering a heated fluid to the chamber to heat the microfluidic device, and a cooling control device in liquid communication with the chamber for delivering a cooled fluid to the chamber to cool the microfluidic device. A temperature control unit in liquid communication with a cooling element and/or a heating element are used to regulate temperature of sequencing substrates and objective lenses for optical detection of sequencing reactions.

Abstract: Methods for high speed, high throughput analysis of polynucleotide sequences, and apparatuses with which to carry out the methods are provided in the invention.

Abstract: The invention relates to methods, systems and devices for mitigation of bubbles in a micro-fluidic environment. For example, the invention relates to methods, systems and devices for mitigation of bubbles from reagents, solvents, formulations and for improving chemical reactions in micro-fluidic systems, such as for fluorescence detection and polynucleotide sequencing.

Abstract: A liquid storage apparatus provides a safe and easy to use device for efficiently managing liquid reagents used in a variety of laboratory equipment. The liquid storage apparatus helps reduce the likelihood of accidents, allows for flexibility of experimental design, and helps maximize the use of chemical regents to prevent waste. The apparatus includes a plurality of containers with a pierceable septum interface at each end. The apparatus also includes a lower array of needles with each of the lower needles in the lower array of needles arranged to penetrate the bottom pierceable septum of a different one of the containers. The apparatus further includes a piercing device arranged to penetrate the top pierceable septum of a different one of the containers. Each of the piercing devices include a passageway so gas can flow into the pierced container.

Abstract: The disclosure provides methods of reducing the range of representation levels of nucleic acid targets. The methods are particularly useful for multi-target analyses benefiting from a low variance of target representations, such as, e.g., single molecule sequencing and/or heterozygous genotyping, and pathogen diagnosis. Two general methods are provided. In Method 1, starting concentrations of probes are adjusted. In Method 2, target-specific probes are “binned,” i.e., several subsets of probes are selected based on similar representation levels. Thereafter, each subset of corresponding targets is extracted, with or without amplification, using a separate portion of the sample (i.e., separate vessels).

Abstract: A method for analyzing genomic data that includes obtaining genomic sequence information from an anonymous individual, processing the information via a secure computerized algorithm, and presenting phenotypic information to the individual based upon the genomic sequence information.

Abstract: The present invention relates to apparatus, systems, and methods for analyzing biological samples. The apparatus, systems, and methods can involve using a vacuum source to pull microfluidic volumes through analytical equipment, such as flow cells and the like. Additionally, the invention involves using optical equipment in conjunction with the analytical equipment to analyze samples and control the operation thereof.

Abstract: The disclosure provides methods and compositions for assessing by sequencing the type and quantity of in vivo or in vitro breakdown products and stability of nucleic acids such as small interfering RNA (siRNA) and other target oligonucleotides. In general, the disclosed methods involve the use of tester oligonucleotides that comprise a double-stranded region, an optional loop (in the case of hairpin testers), and a single-stranded 3? overhang that is complementary to the full-length and a shortened target oligonucleotide. A tester is designed so that the 3? end of a respective target oligonucleotide anneals to the overhang immediately adjacent to the 5? end of the tester. The juxtaposed ends of the tester and target at adjacent positions allow for a ligase to ligate the chain if there is a match between a tester and its respective target.

Abstract: The disclosure provides methods that improve fidelity of the sequencing-by-synthesis reactions, including methods that reduce impurities and contamination in various reagents, reaction mixtures, and other components of the sequencing systems.

Abstract: The disclosure provides methods and compositions for reducing nucleotide impurities in reagents and reaction mixtures. Generally, the methods of the invention involve the inclusion of so-called “scrubbing oligonucleotides” (or “scrubbers”) that preferentially incorporate nucleotide impurities, thereby reducing available free impurities. The disclosure further provides methods of sequencing a target nucleic acid by synthesis that utilize “live” scrubbing. Scrubbing oligonucleotides of various structures are disclosed, including hairpin scrubbers and homopolymeric scrubbers.

Abstract: The disclosure provides methods of generating paired reads in sequencing-by-synthesis process, particularly, in systems with relatively short read lengths (e.g., 15-35 bases), such as for example, in single molecule sequencing by synthesis. Several implementations of the methods are provided. Of particular advantage are the methods that permit re-sequencing of the template, which yields lower error rates. The invention further provides methods of using paired reads, for example, for positioning them over repeats or for assembly into large sequences, including whole genome assembly.

Abstract: The invention provides methods for sequencing a polynucleotide comprising stopping an extension cycle in a sequence by synthesis reaction before the reaction has run to near or fill completion.

Abstract: The invention provides nucleotide analogs for use in sequencing nucleic acid molecules.

Abstract: The invention provides for nucleotide analogs and methods of using the same, e.g., for sequencing nucleic acids.

Abstract: The disclosure provides a method of sequencing a nucleic acid molecule that contains two or more target regions to be sequenced (such as, for example, barcodes). The invention is advantageous for sequencing by synthesis two or more target regions whose combined lengths plus the length of any intermediate sequence exceeds the available read length on a given sequencing platform. The methods of the invention utilize nucleic acid constructs containing at least the following elements: a complement of a first universal primer, a first target sequence, an optional polynucleotide spacer, a complement of a second universal primer, and a second target sequence. A first round of sequencing by synthesis is performed to sequence the first target sequence by elongating the first universal primer. Once the sequence of the first target region is obtained, and before the complement of the second primer is reached, the first round of sequencing is terminated.

Abstract: The disclosure provides methods of capturing target nucleic acids (e.g., gene or gene fragments) onto a solid support for further analysis. The disclosed methods utilize a capture probe that selectively circularizes only the target nucleic acid. Following the circularization of the target, the linear, non-target, nucleic acids are removed from the sample. Next, the circularized target is linearized and bound to a solid support. To allow for linearization, the capture probe may include a cleavage site that can be a noncanonical nucleotide(s) (e.g., uracil in DNA) and/or a rare-cutter site (e.g., the Not I restriction site). In some embodiments, the target nucleic acid is captured onto a support without an intermediate amplification step.

Abstract: A multi-channel flow cell can allow for reduced cross-contamination in sample loading and the ability to observe activity within the flow cell once the channels are loaded. A multi-channel flow cell includes a plurality of independently-addressable channels sandwiched between a two substrates. Each of the channels can be coated with a layer that facilitates support-binding of an analyte. Each of the channels terminates on one end in an inlet and on the other end in an outlet. A loading block having inlet ports that match the inlets of the channels can be mated to the inlets of the channels, and an outlet block can be mated to the outlets of the channels. Analytes can be introduced into the channels via the inlet ports of the loading block and are pulled through the channels by capillary action or by vacuum.

Abstract: The invention provides methods for improving the fidelity of a sequencing-by-synthesis reaction by resequencing at least a portion of a nucleic acid template.