Provided herein are methods of detecting an analyte of interest to interrogate spatial gene expression in a sample using RNA-templated ligation.

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).

The disclosure provides methods and systems for nucleic acid amplification including isothermal nucleic acid amplification.

The disclosure provides methods and systems for nucleic acid amplification including isothermal nucleic acid amplification.

The present invention relates to highly sensitive and specific methods for detection of nucleic acids, which for example are useful for detection of rare mutations, or for detection of low-abundance variants in nucleic acids sequences. The methods involve an asymmetric incremental polymerase reaction (AIPR) followed by an exponential polymerase chain reaction (PCR).

The present invention relates to highly sensitive and specific methods for detection of nucleic acids, which for example are useful for detection of rare mutations, or for detection of low-abundance variants in nucleic acids sequences. The methods involve an asymmetric incremental polymerase reaction (AIPR) followed by an exponential polymerase chain reaction (PCR).

Homogenous detection during or following PCR amplification, preferably LATE-PCR, utilizing fluorescent DNA dye and indirectly excitable labeled primers and probes, improves reproducibility and quantification. Low-temperature homogeneous detection during or following non-symmetric PCR amplification, preferably LATE-PCR, utilizing fluorescent DNA dye and indirectly excitable labeled mismatch-tolerant probes permits analysis of complex targets. Sequencing sample preparation methods following LATE-PCR amplifications reduce complexity and permit “single-tube” processing.

Homogenous detection during or following PCR amplification, preferably LATE-PCR, utilizing fluorescent DNA dye and indirectly excitable labeled primers and probes, improves reproducibility and quantification. Low-temperature homogeneous detection during or following non-symmetric PCR amplification, preferably LATE-PCR, utilizing fluorescent DNA dye and indirectly excitable labeled mismatch-tolerant probes permits analysis of complex targets. Sequencing sample preparation methods following LATE-PCR amplifications reduce complexity and permit “single-tube” processing.

Methods and kits are provided for nucleic acid analysis. In an illustrative method a target nucleic acid is amplified using a first primer and a second primer, wherein the first primer comprises a probe element specific for a locus of the target nucleic acid and a template-specific primer region, and the probe element is 5′ of the template-specific primer region, subsequently allowing the probe element to hybridize to the locus to form a hairpin, generating a melting curve for the probe element by measuring fluorescence from a dsDNA binding dye as the mixture is heated, wherein the dye is not covalently bound to the first primer, and analyzing the shape of the melting curve. Kits may include one or more of the first and second primers, the dsDNA binding dye, a polymerase, and dNTPs.