The invention relates to simplified means of using biological predictive relationships, in some instances reducing the determination of complex gene networks and relative expression patterns to a single reading.

The invention relates to simplified means of using biological predictive relationships, in some instances reducing the determination of complex gene networks and relative expression patterns to a single reading.

Methods for controlling for non-systematic error in an amplification-based next generation sequencing (NGS) library preparation are described, which method includes using an internal amplification control (IAC) sharing identical priming sites to a native nucleic acid target template of interest in a NGS library preparation.

Methods for controlling for non-systematic error in an amplification-based next generation sequencing (NGS) library preparation are described, which method includes using an internal amplification control (IAC) sharing identical priming sites to a native nucleic acid target template of interest in a NGS library preparation.

The invention features compositions and methods that are useful for the measurement of the quantity of a nucleic acid target in a sample.

The invention features compositions and methods that are useful for the measurement of the quantity of a nucleic acid target in a sample.

Disclosed herein is a method of quantifying a mutant allele burden of a target gene in a subject. The method includes providing a first plasmid that includes a mutant allele sequence and an internal control sequence, and a second plasmid that includes a wild-type allele sequence and the internal control sequence, and subjecting DNA of the subject to quantitative polymerase chain reaction to measure a mutant allele expression level of the target gene, so as to determine the mutant allele burden of the target gene in the subject based on a standard curve of the mutant allele burden of the target gene created by serial dilution of the first and second plasmids.

Disclosed herein is a method of quantifying a mutant allele burden of a target gene in a subject. The method includes providing a first plasmid that includes a mutant allele sequence and an internal control sequence, and a second plasmid that includes a wild-type allele sequence and the internal control sequence, and subjecting DNA of the subject to quantitative polymerase chain reaction to measure a mutant allele expression level of the target gene, so as to determine the mutant allele burden of the target gene in the subject based on a standard curve of the mutant allele burden of the target gene created by serial dilution of the first and second plasmids.

A hybridization probe solution containing at least one hybridization probe is applied to final sample handling blank(s) to produce baited final sample handling blank(s), and identical hybridization probe is applied to final control blank(s) carrying transfer substrate identical to that applied to the sample handling blank(s) but isolated from the sample biological materials, to thereby produce at least one baited final control blank. The baited final sample handling blank(s) and baited final control blank(s) are fed into a DNA sequencer to sequence sample bait-captured DNA carried by the baited final sample handling blank and control bait-captured DNA carried by the baited final control blank, respectively. The sample bait-captured DNA is compared to the control bait-captured DNA and genetic components that are common to the final sample handling blank and the final control blank and pass a statistical significance test are discounted from a final identified genetic sequence.

A hybridization probe solution containing at least one hybridization probe is applied to final sample handling blank(s) to produce baited final sample handling blank(s), and identical hybridization probe is applied to final control blank(s) carrying transfer substrate identical to that applied to the sample handling blank(s) but isolated from the sample biological materials, to thereby produce at least one baited final control blank. The baited final sample handling blank(s) and baited final control blank(s) are fed into a DNA sequencer to sequence sample bait-captured DNA carried by the baited final sample handling blank and control bait-captured DNA carried by the baited final control blank, respectively. The sample bait-captured DNA is compared to the control bait-captured DNA and genetic components that are common to the final sample handling blank and the final control blank and pass a statistical significance test are discounted from a final identified genetic sequence.