Provided herein are compositions and methods for accurate and scalable Primary Template-Directed Amplification (PTA) nucleic acid amplification and sequencing methods, and their applications for research, diagnostics, and treatment.

A LAMP primer set includes original LAMP primers of FIP, BIP, F3, and B3, and at least one autonomy primer. The original LAMP primers target regions F3, F2, F1C, B1C, B2, and B3 on nucleic acids, and the regions F3, F2, F1, B1C, B2C and B 3 C are located in order from 5′ end to 3′ end of a forward strand of the nucleic acids. The primer FIP includes oligonucleotides targeting F1C and F2, and the primer BIP includes oligonucleotides targeting B1C and B2. The at least one autonomy primer targets a region located beyond a region from F3 to B3.

An assay is provided for the rapid detection of nucleic acids via a modified LAMP reaction coupled with colorimetric reporter utilizing a gold nanoparticle—peptide nucleic acid (AuNP-PNA) probe system.

An assay is provided for the rapid detection of nucleic acids via a modified LAMP reaction coupled with colorimetric reporter utilizing a gold nanoparticle—peptide nucleic acid (AuNP-PNA) probe system.

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.

The present invention features compositions and methods for amplifying a target oligonucleotide in a sample, including detection of the target oligonucleotide in real time.

The present invention features compositions and methods for amplifying a target oligonucleotide in a sample, including detection of the target oligonucleotide in real time.

Provided herein are compositions and methods for accurate and scalable Primary Template-Directed Amplification (PTA) nucleic acid amplification and sequencing methods, and their applications for mutational analysis in research, diagnostics, and treatment. Further provided herein are multiomics methods for parallel analysis of DNA, RNA, and/or proteins from single cells. Provided herein are methods of multiomic single-cell analysis comprising: (a) isolating a single cell from a population of cells; (b) sequencing a cDNA library comprising polynucleotides amplified from mRNA transcripts from the single cell; and (c) sequencing a genome of the single cell.

Provided herein are compositions and methods for accurate and scalable Primary Template-Directed Amplification (PTA) nucleic acid amplification and sequencing methods, and their applications for mutational analysis in research, diagnostics, and treatment. Further provided herein are multiomics methods for parallel analysis of DNA, RNA, and/or proteins from single cells. Provided herein are methods of multiomic single-cell analysis comprising: (a) isolating a single cell from a population of cells; (b) sequencing a cDNA library comprising polynucleotides amplified from mRNA transcripts from the single cell; and (c) sequencing a genome of the single cell.