The present invention relates to a method for detecting nucleic acid (NA) molecules in samples. More particularly, the present invention relates to an improved digital PCR-based method for detecting specific nucleic acid sequence(s). The present invention is useful for research and diagnostic applications with increased sensitivity and accuracy. The present invention also provides a kit for performing the method for assessingnucleic acids in samples as described herein.

The present invention relates to a method for detecting nucleic acid (NA) molecules in samples. More particularly, the present invention relates to an improved digital PCR-based method for detecting specific nucleic acid sequence(s). The present invention is useful for research and diagnostic applications with increased sensitivity and accuracy. The present invention also provides a kit for performing the method for assessingnucleic acids in samples as described herein.

Provided herein are methods of determining a surgical margin and the site and size of a tissue to be resected from a subject, and methods of use thereof.

Provided herein are methods of determining a surgical margin and the site and size of a tissue to be resected from a subject, and methods of use thereof.

The invention relates to methods of tagging analytes in a sample.

A method for spatially tagging nucleic acids of a biological specimen, including steps of (a) providing a solid support comprising different nucleic acid probes that are randomly located on the solid support, wherein the different nucleic acid probes each includes a barcode sequence that differs from the barcode sequence of other randomly located probes on the solid support; (b) performing a nucleic acid detection reaction on the solid support to locate the barcode sequences on the solid support; (c) contacting a biological specimen with the solid support that has the randomly located probes; (d) hybridizing the randomly located probes to target nucleic acids from portions of the biological specimen; and (e) modifying the randomly located probes that are hybridized to the target nucleic acids, thereby producing modified probes that include the barcode sequences and a target specific modification, thereby spatially tagging the nucleic acids of the biological specimen.

A method for spatially tagging nucleic acids of a biological specimen, including steps of (a) providing a solid support comprising different nucleic acid probes that are randomly located on the solid support, wherein the different nucleic acid probes each includes a barcode sequence that differs from the barcode sequence of other randomly located probes on the solid support; (b) performing a nucleic acid detection reaction on the solid support to locate the barcode sequences on the solid support; (c) contacting a biological specimen with the solid support that has the randomly located probes; (d) hybridizing the randomly located probes to target nucleic acids from portions of the biological specimen; and (e) modifying the randomly located probes that are hybridized to the target nucleic acids, thereby producing modified probes that include the barcode sequences and a target specific modification, thereby spatially tagging the nucleic acids of the biological specimen.

Embodiments of the disclosure encompass highly sensitive and quantitative methods for single-cell sequencing of total RNA. In particular cases, methods utilize annealing of multiple primers to RNA, polytailing of single stranded DNA reverse transcribed therefrom, and utilization of bar codes in primers for amplification of amplicons produced from second strand synthesis.

Embodiments of the disclosure encompass highly sensitive and quantitative methods for single-cell sequencing of total RNA. In particular cases, methods utilize annealing of multiple primers to RNA, polytailing of single stranded DNA reverse transcribed therefrom, and utilization of bar codes in primers for amplification of amplicons produced from second strand synthesis.

Described herein are methods for determining a sequence of a region of interest from an mRNA molecule. Sequenced polynucleotides can include a barcode region, a homopolymer region (e.g., a poly-A region), and a target region associated with the mRNA molecule. According to some methods, the barcode region omits the same base present in the homopolymer region. According to some methods, extension of the primer used for sequencing is stalled within the homopolymer region. According to some methods, sequencing flow cycles and the different barcode regions of the polynucleotides configured are such that the primer is extended to the end of the barcode region across the plurality of polynucleotides before being extended into the homopolymer region. According to some methods, two primers or a cleavable primer is used to separately sequence the barcode region and the target region.