A sequencing system includes an automated sequencing instrument adapted to determine variant calls for one or more extracted polynucleotide samples with a performance of at least 98.5% raw read accuracy and a total instrument run time in a range of 16 hours to 22 hours to determine variant calls when sequencing 6 cfTNA extracted polynucleotide samples using a targeted assay with one nucleic acid pool per sample.

Provided herein are methods for mapping modified nucleotide residues in nucleic acids. The methods include providing a nucleic acid sample in which non-target or target modified and unmodified nucleotide residues are converted to form of a different nucleotide (such a “C” being converted to “T”). Second strand synthesis is then performed on the converted nucleic acids using a set of anchored-base primers. Each primer in the set of anchored-base primers comprises one or more anchor bases at the 3′ terminus that are complementary to the target nucleotide (e.g., “G” or “CpG”), and a sequence of nucleotides selected from a set of sequences that could be a fully or partially degenerate set of sequences. For example, the sequence could be 5′-XnG-3′ and/or 5′-X(n−1)CG-3′, wherein X is any base, and n=2 to 25. Double-stranded nucleic acid products can be analyzed, for example by amplification and high throughput sequencing.

Provided herein are methods for mapping modified nucleotide residues in nucleic acids. The methods include providing a nucleic acid sample in which non-target or target modified and unmodified nucleotide residues are converted to form of a different nucleotide (such a “C” being converted to “T”). Second strand synthesis is then performed on the converted nucleic acids using a set of anchored-base primers. Each primer in the set of anchored-base primers comprises one or more anchor bases at the 3′ terminus that are complementary to the target nucleotide (e.g., “G” or “CpG”), and a sequence of nucleotides selected from a set of sequences that could be a fully or partially degenerate set of sequences. For example, the sequence could be 5′-XnG-3′ and/or 5′-X(n−1)CG-3′, wherein X is any base, and n=2 to 25. Double-stranded nucleic acid products can be analyzed, for example by amplification and high throughput sequencing.

The invention relates to methods of detecting a genetic variation in a genetic sample from a subject using labeled probes and counting the number of labels in the probes.

The invention relates to methods of detecting a genetic variation in a genetic sample from a subject using labeled probes and counting the number of labels in the probes.

Provided are methods for determining if a cannabis sample comprises hemp or marijuana, or Cannabis sativa and/or Cannabis indica as well as primers and kits for use in the methods.

The description provides two-stage methods of nucleic acid amplification and detection reactions, which are useful for rapid pathogen detection or disease diagnosis. In particular, the description provides a method comprising a first-stage slow rate amplification reaction followed by a plurality of second-stage fast rate amplification reactions that are simultaneously monitored in real-time, and wherein a rapid rate of amplification is indicative of the presence of a site of interest.

The description provides two-stage methods of nucleic acid amplification and detection reactions, which are useful for rapid pathogen detection or disease diagnosis. In particular, the description provides a method comprising a first-stage slow rate amplification reaction followed by a plurality of second-stage fast rate amplification reactions that are simultaneously monitored in real-time, and wherein a rapid rate of amplification is indicative of the presence of a site of interest.

A method for separating a target allele from a mixture of nucleic acids by (a) providing a mixture of nucleic acids in fluidic contact with a stabilized ternary complex that is attached to a solid support, wherein the stabilized ternary complex includes a polymerase, primed nucleic acid template, and next correct nucleotide, wherein the template has a target allele, wherein the next correct nucleotide is a cognate nucleotide for the target allele, and wherein the stabilized ternary complex is attached to the solid support via a linkage between the polymerase and the solid support or via a linkage between the next correct nucleotide and the solid support; and (b) separating the solid support from the mixture of nucleic acids, thereby separating the target allele from the mixture of nucleic acids.

A method for separating a target allele from a mixture of nucleic acids by (a) providing a mixture of nucleic acids in fluidic contact with a stabilized ternary complex that is attached to a solid support, wherein the stabilized ternary complex includes a polymerase, primed nucleic acid template, and next correct nucleotide, wherein the template has a target allele, wherein the next correct nucleotide is a cognate nucleotide for the target allele, and wherein the stabilized ternary complex is attached to the solid support via a linkage between the polymerase and the solid support or via a linkage between the next correct nucleotide and the solid support; and (b) separating the solid support from the mixture of nucleic acids, thereby separating the target allele from the mixture of nucleic acids.