Methods of repairing a partially double-stranded DNA fragment are provided. In some embodiments, the methods comprise (a) contacting the partially double-stranded DNA fragment with one or more primers of a primer population, wherein the partially double-stranded DNA fragment comprises a 3′ overhang and the primer population comprises a random target-hybridizing sequence; (b) extending one or more primers of the primer population along the DNA fragment using a DNA polymerase, thereby producing one or more extended primers annealed to the DNA fragment; and (c) ligating the 3′ end of one or more extended primers to the 5′ end of an extended primer or a strand of the partially double-stranded DNA fragment, thereby providing a repaired DNA fragment.

A primer composition, a kit and a method for detecting microhaplotype loci based on next generation sequencing technology and applications thereof are provided, relating to the technical field of forensic medicine, which are used to amplify 163 microhaplotype loci on human genome. The primer composition includes one or more pairs of primers with sequences as shown in SEQ ID NO: 1˜326. The primer composition involves 163 microhaplotype loci covering 22 autosomes, which can provide more new genetic information in Asian population than the system constructed in the past. In addition, compared with the next generation sequencing kit of STR loci, the kit has better mixture detection capability. Moreover, the microhaplotype genetic markers have high ancestry information content and can distinguish populations in Africa, Europe, South Asia, and East Asia. Therefore, the microhaplotype genetic markers can also be used for ancestry inference in addition to individual identification and parentage testing.

A primer composition, a kit and a method for detecting microhaplotype loci based on next generation sequencing technology and applications thereof are provided, relating to the technical field of forensic medicine, which are used to amplify 163 microhaplotype loci on human genome. The primer composition includes one or more pairs of primers with sequences as shown in SEQ ID NO: 1˜326. The primer composition involves 163 microhaplotype loci covering 22 autosomes, which can provide more new genetic information in Asian population than the system constructed in the past. In addition, compared with the next generation sequencing kit of STR loci, the kit has better mixture detection capability. Moreover, the microhaplotype genetic markers have high ancestry information content and can distinguish populations in Africa, Europe, South Asia, and East Asia. Therefore, the microhaplotype genetic markers can also be used for ancestry inference in addition to individual identification and parentage testing.

Provided herein are compositions and methods for simultaneously measuring target oligonucleotides and protein in single cells. Compositions comprise an antibody-tagged oligonucleotide, including an origin specific barcode handle sequence, a first primer handle sequence, a second primer handle sequence, and a target binding region. The composition may also include an adapter sequence, a unique molecular identifier (UMI), and a poly-A sequence. Methods for simultaneously measuring target oligonucleotides and protein in single cells generally involve delivering a mixture of the composition to a population of cells and encapsulating individual cells in an individual discrete volume comprising PCR primers on a bead. The individual discrete volume may be suspended in a reverse transcription mixture and the nucleotide sequence of the origin specific barcode handle sequence may be detected, thereby assigning the target oligonucleotide and protein of interest to a specific individual discrete volume, while maintaining information about sample origin of the target oligonucleotide.

Provided herein are compositions and methods for simultaneously measuring target oligonucleotides and protein in single cells. Compositions comprise an antibody-tagged oligonucleotide, including an origin specific barcode handle sequence, a first primer handle sequence, a second primer handle sequence, and a target binding region. The composition may also include an adapter sequence, a unique molecular identifier (UMI), and a poly-A sequence. Methods for simultaneously measuring target oligonucleotides and protein in single cells generally involve delivering a mixture of the composition to a population of cells and encapsulating individual cells in an individual discrete volume comprising PCR primers on a bead. The individual discrete volume may be suspended in a reverse transcription mixture and the nucleotide sequence of the origin specific barcode handle sequence may be detected, thereby assigning the target oligonucleotide and protein of interest to a specific individual discrete volume, while maintaining information about sample origin of the target oligonucleotide.

Disclosed herein, inter alia, are polynucleotides, supports, kits, and methods of use thereof for amplifying, immobilizing, and sequencing polynucleotides.

Disclosed herein, inter alia, are polynucleotides, supports, kits, and methods of use thereof for amplifying, immobilizing, and sequencing polynucleotides.

The invention pertains to construction of next-generation DNA sequencing (NGS) libraries for whole genome sequencing, targeted resequencing, sequencing-based screening assays, metagenomics, or any other application requiring sample preparation for NGS.

The invention pertains to construction of next-generation DNA sequencing (NGS) libraries for whole genome sequencing, targeted resequencing, sequencing-based screening assays, metagenomics, or any other application requiring sample preparation for NGS.

A method for specifically and efficiently quantifying the expression of targeted RNA variants with specific terminal sequences suitable to identify multiple isoforms bearing complex heterogeneity in terminal sequences by hybridizing a 5′-Dbs-adapter to the 5′-end of target RNAs, wherein the 5′-Dbs-adapter has a stem-loop structure whose protruding 5′-end base-pairs with the 5′-end of target RNAs, and wherein the loop region of 5′-Dbs-adapter contains a base-lacking spacer which will terminate reverse transcription in a subsequent step; hybridizing a 3′Db-adapter to the 3′-end of target RNAs, wherein the 3′-Db-adapter has a stem-loop structure whose protruding 3′-end base-pairs with the 3′-end of target RNAs; ligating both adapters with target RNAs by RN12 ligation to form a “dumbbell-like” structure; and, amplifying and quantifying the ligation product by RT-PCR.