Size standards for next-generation sequencing
AssigneeRegents of the University of Minnesota
InventorsDaryl M. Gohl, Kenneth B. Beckman
Life of this patent
Estimating the term…
Abstract
Provided herein are synthetic size standards that allow one to detect size bias in a sample that includes a plurality of polynucleotides. The size standards can provide an internal control to detect and correct for size bias in data obtained after manipulating and/or processing of sample polynucleotide. Also provided herein are methods for detecting size bias in a sample or in a sequencing run.
Claims13
A composition comprising a plurality of synthetic size standard polynucleotides, each synthetic size standard polynucleotide of the plurality comprising one or more next-generation sequencing adapter-flanked barcodes, wherein the plurality comprises defined ratios of synthetic size standard polynucleotides of at least two defined lengths, the plurality defining a continuous or random length distribution spanning a defined portion of a genome.
A method for detecting size bias in a sample comprising one or more template polynucleotides, the method comprising
- obtaining a sample comprising one or more template polynucleotides;
- spiking the sample with a plurality of synthetic size standard polynucleotides designed to detect size bias between two template polynucleotides of the sample, each synthetic size standard polynucleotide of the plurality comprising a next-generation sequencing adapter-flanked size barcode, wherein the plurality comprises defined ratios of synthetic size standard polynucleotides of multiple defined lengths;
- sequencing at least a portion of each template polynucleotide of the spiked sample using a sequencer corresponding to the next-generation sequencing adapter;
- measuring the frequency of occurrence of the size barcode;
- and
- comparing the measured occurrence of the size barcodes to an expected frequency occurrence of the size barcodes, thereby generating a size standard polynucleotide size bias value.
Dependencies are inferred from the claim text.