Life Technologies Corporation

Applied Biosystems, LLC

Applera Corporation

David C Woo

Jeffrey A Marks

Thomas Wessel

Yong Chu

Stephanie Jo Schneider

Jacob K Freudenthal

Manjula Aliminati

Sylvia Chang

Clinton Lewis

Nasser M Abbasi

David Denny

Siva Kumar Samsani

Yoke Peng Lim

Marcin Sikora

Nivedita Sumi Majumdar

Gordon A Janaway

Stephen J Gunstream

Yerramalli Subramaniam

Alan Russell Stanford

Joanna Lankester

Daniel Wessel

John David Morgenthaler

Wallace George

John P Bodeau

Mark A McCoy

Rylan Schaeffer

Shweta Raizada

Mark A Mc Coy

A method of automatically sequencing or basecalling one or more DNA (deoxyribonucleic acid) molecules of a biological sample is described. The method comprises using a capillary electrophoresis genetic analyzer to measure the biological sample to obtain at least one input trace comprising digital data corresponding to fluorescence values for a plurality of scans. Scan labelling probabilities for the plurality of scans are generated using a trained artificial neural network comprising a plurality of layers including convolutional layers. A basecall sequence comprising a plurality of basecalls for the one or more DNA molecules based on the scan labelling probabilities for the plurality of scans is determined.

Basecaller with dilated convolutional neural network

A computer-implemented method for designing a digital PCR (dPCR) experiment is provided. The method includes receiving, from a user, a selection of optimization type. The optimization type may be maximizing the dynamic range, minimizing the number of substrates including reaction sites needed for the experiment, determining a dilution factor, or determining the lower limit of detection, for example. The method further includes receiving, from the user, a precision measure for an experiment, and a minimum concentration of a target in a reaction site for the experiment. The method also includes determining a set of dPCR experiment design factors for the experiment based on the optimization type. The set of dPCR experiment design factors is then displayed to the user.

Methods and systems for a digital PCR experiment designer

A biological analysis system and an associated method are provided. The method is used for recovering off scale data in an image produced by a camera in a capillary electrophoresis instrument. The method comprises the steps of identifying bins of the image where electron counts exceed a maximum number of counts; setting an off-scale flag for the identified bins; and processing the image to obtain a recovered dye signal, based on the flag set for each bin, and using a dye matrix.

Methods and apparatus for extended dynamic range from single exposures in capillary electrophoresis

In one exemplary embodiment, a method for validating an instrument is provided. The method includes receiving amplification data from a validation plate to generate a plurality of amplification curves. The validation plate includes a sample of a first quantity and a second quantity, and each amplification curve includes an exponential region. The method further includes determining a set of fluorescence thresholds based on the exponential regions of the plurality of amplification curves and determining, for each fluorescence threshold of the set, a first set of cycle threshold (C) values of amplification curves generated from the samples of the first quantity and a second set of Cvalues of amplification curves generated from the samples of the second quantity. The method includes calculating if the first and second quantities are sufficiently distinguishable based on Cvalues at each of the plurality of fluorescence thresholds.

Methods and systems for instrument validation

In one exemplary embodiment, a method for detecting variants in electropherogram data is provided. The method includes receiving electropherogram data from an instrument and analyzing the electropherogram data to identify mixed bases in the electropherogram data. The method further includes validating the identified mixed bases. Then the method includes determining variants in the electropherogram data based on the validated mixed bases.

Methods and systems for variant detection

In one exemplary embodiment, a method for detecting variants in electropherogram data is provided. The method includes receiving electropherogram data from an instrument and analyzing the electropherogram data to identify mixed bases in the electropherogram data. The method further includes identifying features within the electropherogram data indicative of errors and validating the identified mixed bases. Then the method includes determining variants in the electropherogram data based on the validated mixed bases.

In one exemplary embodiment, a method for calibrating an instrument is provided. The instrument includes an optical system capable of imaging florescence emission from a plurality of reaction sites. The method includes performing a region-of-interest (ROI) calibration to determine reaction site positions in an image. The method further includes performing a pure dye calibration to determine the contribution of a fluorescent dye used in each reaction site by comparing a raw spectrum of the fluorescent dye to a pure spectrum calibration data of the fluorescent dye. The method further includes performing an instrument normalization calibration to determine a filter normalization factor. The method includes performing an RNase P validation to validate the instrument is capable of distinguishing between two different quantities of sample.

Methods and systems for biological instrument calibration

The invention discloses a system and methods for quantitating the presence of nucleic acid sequences by evaluation of amplification data generated using real-time PCR. In one aspect, the methods may be adapted to identify a threshold and threshold cycle for one or more reactions based upon evaluation of exponential and baseline regions for each amplification reaction. The methodology used in the analysis may be readily automated such that subjective user interpretation of the data is substantially reduced or eliminated.

Automatic threshold setting and baseline determination for real-time PCR

Systems and methods are provided that comprise calibration techniques and associated systems that identify the two-dimensional position, or other alignment or positioning, of sample wells or other calibration objects located in a sample well plate, or other surface or area of interest. In some embodiments, calibration of the plate and/or positioning and/or alignment with respect to detection optics can be performed in multiple stages for two or more dimensions.

Calibrating the positions of a rotating and translating two-dimensional scanner

Methods and systems for processing signals to minimize the effects of dye crosstalk. Deconvolution of multiplexed dye signals can include corrections for residual error determined from experimental measurements. Residual error corrections can account for reaction or assay specific factors and modify the subsequent filtering of signals. Correction values can be determined for specific dye-probe conjugates to minimize dye crosstalk and may be combined with residual error correction to further minimize errors in spectral deconvolution. Apparatus, systems, and computer-readable media can process signals and modify filters based on values obtained using the methods.

Minimizing effects of dye crosstalk

Aspects of the present invention describe a method and apparatus for automating quality control for gene expression data. A computer based device receives gene expression data associated with a spectral species and genetic sample in each well of a plate. Gene expression data may be received from a sequence detection instrument performing one or more gene expression related operations for each of the wells of the plate. The computer based device identifies gene expression data determined to have anomalous characteristics according to a set of one or more quality control metrics and may conditionally flag one or more wells of the plate affected by the anomalous characteristics. Filters can then be selectively applied to temporarily or permanently remove the flagged data from subsequent gene expression studies.

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Foster City, CA, United States of America

David C Woo