In some embodiments, methods for ligating nucleic acid ends comprise: conducting a nucleic acid ligation reaction in the presence of at least one agent that generates a ligatable terminal 5′ phosphate group by removing an adenylate group from a terminal 5′ phosphate of a nucleic acid. In some embodiments, an aprataxin enzyme can catalyze removal of an adenylate group from a terminal 5′ phosphate of a nucleic acid. In some embodiments, methods for ligating nucleic acid ends comprise: conducting a nucleic acid ligation reaction in the presence of an aprataxin enzyme under conditions suitable for ligating nucleic acid ends.

In some embodiments, methods for ligating nucleic acid ends comprise: conducting a nucleic acid ligation reaction in the presence of at least one agent that generates a ligatable terminal 5′ phosphate group by removing an adenylate group from a terminal 5′ phosphate of a nucleic acid. In some embodiments, an aprataxin enzyme can catalyze removal of an adenylate group from a terminal 5′ phosphate of a nucleic acid. In some embodiments, methods for ligating nucleic acid ends comprise: conducting a nucleic acid ligation reaction in the presence of an aprataxin enzyme under conditions suitable for ligating nucleic acid ends.

A device for extracting a nucleic acid from a sample liquid includes a heating element configured to be connected to an extraction nucleic acid. The extraction nucleic acid is at least partly complementary to the nucleic acid to be extracted from the sample liquid. The heating element is heatable to a temperature that is equal to or higher than a denaturing temperature of the nucleic acid bound to the extraction nucleic acid.

A device for extracting a nucleic acid from a sample liquid includes a heating element configured to be connected to an extraction nucleic acid. The extraction nucleic acid is at least partly complementary to the nucleic acid to be extracted from the sample liquid. The heating element is heatable to a temperature that is equal to or higher than a denaturing temperature of the nucleic acid bound to the extraction nucleic acid.

The present invention discloses a convective PCR apparatus by using a transparent conductive thin film to replace the traditional metal heater. The PCR reaction is activated when the container with reagents contacted the heated transparent conductive thin film and the temperature inside the container raised to initiate the convective circulation. Also, the present invention could apply for a quantitative PCR reaction by adding a specific probe, a fluorescent dye, a light source, or a photon receiver.

The present invention provides an application of a cold-active bacteriophage protein in a room temperature nucleic acid amplification reaction; the cold-active bacteriophage is selected from vB_EcoM-VR5, vB_EcoM-VR7, and vB_EcoM-VR20,vB_EcoM-VR25, or vB_EcoM-VR26, and the cold-active bacteriophage protein is a uvsX protein, a uvsY protein and a gp32 protein and/or a variant protein having corresponding functions. Preferably, the uvsX protein and the variant protein thereof are selected from any sequence of SEQ ID Nos. 1-23 or 30, the uvsY protein and the variant protein thereof are selected from any sequence of SEQ ID Nos.27-29 or 32, and the gp32 protein and the variant protein thereof are selected from any sequence of SEQ ID Nos.24-26 or 31. The present invention further provides a room temperature nucleic acid amplification reaction system containing the cold-active bacteriophage protein.

The present invention provides an application of a cold-active bacteriophage protein in a room temperature nucleic acid amplification reaction; the cold-active bacteriophage is selected from vB_EcoM-VR5, vB_EcoM-VR7, and vB_EcoM-VR20,vB_EcoM-VR25, or vB_EcoM-VR26, and the cold-active bacteriophage protein is a uvsX protein, a uvsY protein and a gp32 protein and/or a variant protein having corresponding functions. Preferably, the uvsX protein and the variant protein thereof are selected from any sequence of SEQ ID Nos. 1-23 or 30, the uvsY protein and the variant protein thereof are selected from any sequence of SEQ ID Nos.27-29 or 32, and the gp32 protein and the variant protein thereof are selected from any sequence of SEQ ID Nos.24-26 or 31. The present invention further provides a room temperature nucleic acid amplification reaction system containing the cold-active bacteriophage protein.

The present invention relates to a method for determining presence of a pre-determined nucleic acid sequence in a sample, the method comprising the steps of adding one or more enzyme(s) providing activities of RNA- and/or DNA-dependent DNA polymerase activity and strand-displacement activity to the sample to be analysed for the presence of the pre-determined nucleic acid sequence; adding at least five DNA primers to the sample to be analysed for the presence of the pre-determined nucleic acid sequence, wherein at least one DNA primer comprises a sequence hybridisable to the nucleic acid sequence and at least one DNA primer comprises a sequence hybridisable to the DNA sequence reverse-complementary to the nucleic acid sequence; incubating the sample resulting at a fixed temperature; determining whether an elongated DNA sequence is present in the sample, wherein presence of the elongated DNA sequence in the sample is indicative of the presence of the pre-determined nucleic acid sequence in the sample, wherein the sample is obtained from an animal and wherein no F3 primer is used.

The present invention relates to a method for determining presence of a pre-determined nucleic acid sequence in a sample, the method comprising the steps of adding one or more enzyme(s) providing activities of RNA- and/or DNA-dependent DNA polymerase activity and strand-displacement activity to the sample to be analysed for the presence of the pre-determined nucleic acid sequence; adding at least five DNA primers to the sample to be analysed for the presence of the pre-determined nucleic acid sequence, wherein at least one DNA primer comprises a sequence hybridisable to the nucleic acid sequence and at least one DNA primer comprises a sequence hybridisable to the DNA sequence reverse-complementary to the nucleic acid sequence; incubating the sample resulting at a fixed temperature; determining whether an elongated DNA sequence is present in the sample, wherein presence of the elongated DNA sequence in the sample is indicative of the presence of the pre-determined nucleic acid sequence in the sample, wherein the sample is obtained from an animal and wherein no F3 primer is used.

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.