A reaction vessel comprises a lower chamber with a first volume, and an upper chamber with a second volume greater than the first volume. A thermocycling system for heating the reaction vessel includes a lower heating zone to heat the lower chamber, an upper heating zone to heat the upper chamber, and a lid heater to heat an opening of the upper chamber. A method comprises loading a sample into a lower chamber of a reaction vessel, thermocycling the lower chamber using a lower heating zone of the thermo cycler, combining an additive into the sample to produce a combination filling the lower chamber and at least partially filling an upper chamber of the reaction vessel, and incubating the upper and lower chambers using the lower heating zone and an upper heating zone. The lower and upper chambers can have different wall thicknesses to facilitate heat transfer.

The present invention provides for stable nucleotide reagents used for nucleic acid amplification by PCR and RT-PCR (Reverse Transcriptase-PCR) that comprises modified nucleoside polyphosphates. The present invention also provides for methods for using the modified nucleoside polyphosphates for detecting the presence or absence of a target nucleic acid sequence in a sample in an amplification reaction.

The present invention provides for stable nucleotide reagents used for nucleic acid amplification by PCR and RT-PCR (Reverse Transcriptase-PCR) that comprises modified nucleoside polyphosphates. The present invention also provides for methods for using the modified nucleoside polyphosphates for detecting the presence or absence of a target nucleic acid sequence in a sample in an amplification reaction.

The present disclosure provides methods of preparing a fixed biological sample for use in an assay, wherein the method includes treatment of the sample with a low temperature active protease, optionally, in combination with an un-fixing reagent. The disclosure also provides assay methods, include partition-based methods, for fixed biological sample that use the low temperature protease treatment in combination with an un-fixing reagent. Kits comprising protease compositions, un-fixing agent compositions, and other assay reagents for use in the methods are also provided.

The present disclosure provides methods of preparing a fixed biological sample for use in an assay, wherein the method includes treatment of the sample with a low temperature active protease, optionally, in combination with an un-fixing reagent. The disclosure also provides assay methods, include partition-based methods, for fixed biological sample that use the low temperature protease treatment in combination with an un-fixing reagent. Kits comprising protease compositions, un-fixing agent compositions, and other assay reagents for use in the methods are also provided.

The present disclosure is drawn to compositions, methods, and systems for loop-mediated isothermal amplification (LAMP) analysis on a solid phase medium. The composition can comprise one or more target primers, a DNA polymerase, and a re-solubilization agent. The composition can be substantially free of non-pH sensitive agents capable of discoloring the solid phase medium. The method can comprise providing an assembly of a solid phase medium, depositing a biological sample onto the solid phase medium, and heating the assembly to an isothermal temperature sufficient to facilitate a LAMP reaction. The system can comprise a composition and a solid phase medium on to which the composition is deposited.

The present disclosure is drawn to compositions, methods, and systems for loop-mediated isothermal amplification (LAMP) analysis on a solid phase medium. The composition can comprise one or more target primers, a DNA polymerase, and a re-solubilization agent. The composition can be substantially free of non-pH sensitive agents capable of discoloring the solid phase medium. The method can comprise providing an assembly of a solid phase medium, depositing a biological sample onto the solid phase medium, and heating the assembly to an isothermal temperature sufficient to facilitate a LAMP reaction. The system can comprise a composition and a solid phase medium on to which the composition is deposited.

The present disclosure relates to the field of biotechnology, in particular, to a combination product for detecting DNA. The product includes ribonuclease H II and a probe; the probe is a single-stranded probe and has a sequence which can be partially or entirely complementary to a target DNA molecule to be detected, and one or more RNA bases are embedded in the complementary region of the probe and divide the complementary region of the probe into at least two segments, each of which independently has DNA bases and base substitutions of less than or equal to 13 in total.

The present disclosure relates to the field of biotechnology, in particular, to a combination product for detecting DNA. The product includes ribonuclease H II and a probe; the probe is a single-stranded probe and has a sequence which can be partially or entirely complementary to a target DNA molecule to be detected, and one or more RNA bases are embedded in the complementary region of the probe and divide the complementary region of the probe into at least two segments, each of which independently has DNA bases and base substitutions of less than or equal to 13 in total.

The present disclosure generally relates to systems and methods for detecting viruses, e.g., using microfluidic devices. Certain embodiments are generally directed to systems and methods that are able to detect pathogens such as viruses or bacteria by encapsulating a sample in droplets, and applying amplification reagents to the droplets able to amplify nucleic acids therein, e.g., using loop mediated isothermal amplification (LAMP) or other amplification techniques. In addition, some aspects are generally directed to identifying a species in a sample, e.g., at very low concentrations. In some cases, the sample may be broken into droplets, arid the droplets determined to determine the species.