The invention generally relates to methods for increasing the amount of DNA available for analysis when using partitioned samples and parallel processing. For example, double-stranded DNA can be dissociated into two single-stranded components, and the single strands partitioned into different droplets prior to analysis. The disclosed methods are useful for performing digital PCR analysis on samples where the target DNA is not in abundance, for example when the sample originates from a body fluid or an FFPE sample.

The present invention is generally drawn to a novel method for determining a SNP (single nucleotide polymorphism) genotype using a PTO-SNV (Probing and Tagging Oligonucleotide for Single Nucleotide Variation). The present invention provides novel protocols for SNP genotyping in which only one allele-specific oligonucleotide permits in a SNP genotyping reaction to determine whether a target nucleic acid sequence to be analyzed is homozygous or heterozygous for the SNP allele of interest or has no SNP allele of interest.

The present invention is generally drawn to a novel method for determining a SNP (single nucleotide polymorphism) genotype using a PTO-SNV (Probing and Tagging Oligonucleotide for Single Nucleotide Variation). The present invention provides novel protocols for SNP genotyping in which only one allele-specific oligonucleotide permits in a SNP genotyping reaction to determine whether a target nucleic acid sequence to be analyzed is homozygous or heterozygous for the SNP allele of interest or has no SNP allele of interest.

Provided herein are reagents and kits for detection of multiple target sequences in a single-tube, single-color assay, and methods of use thereof. In particular, multiplex assays are provided for the detection of Mycobacterium tuberculosis complex target sequences (e.g., katG, rpoB, inhA promotor, pncA, etc.).

Provided herein are reagents and kits for detection of multiple target sequences in a single-tube, single-color assay, and methods of use thereof. In particular, multiplex assays are provided for the detection of Mycobacterium tuberculosis complex target sequences (e.g., katG, rpoB, inhA promotor, pncA, etc.).

Methods and reagent for determining the presence and/or for quantifying the amount of a target nucleic acid sequences in a sample are provided. In some aspects, the methods comprise performing a melt analysis by detecting, a signal from a probe at a temperature that is lower than the Tm of the probe and a signal at a temperature that is higher than the Tm of the probe, without detecting a signal at the Tm of the probe.

Methods and reagent for determining the presence and/or for quantifying the amount of a target nucleic acid sequences in a sample are provided. In some aspects, the methods comprise performing a melt analysis by detecting, a signal from a probe at a temperature that is lower than the Tm of the probe and a signal at a temperature that is higher than the Tm of the probe, without detecting a signal at the Tm of the probe.

Disclosed herein is a nucleic acid amplification process referred to as Linear-Expo-Linear Polymerase Chain Reaction (LEL-PCR).

Disclosed herein is a nucleic acid amplification process referred to as Linear-Expo-Linear Polymerase Chain Reaction (LEL-PCR).

In certain aspects, the invention disclosed herein relates to the isothermal amplification of probe linkage products to generate specific amplified signals. In some aspects, the invention provides methods, reagents, and kits for carrying out such amplification via the isothermal chain reaction (ICR).