Methods of detecting methylated CpG are provided. Accordingly, there is provided a method of determining CpG methylation status in a DNA sample, the method comprising: (a) subjecting the DNA sample to bisulfite conversion; (b) amplifying said DNA sample following said (a) to obtain an amplified DNA sample; (c) labeling CpG sites in said amplified DNA sample with a label to obtain a labeled DNA sample; (d) contacting said labeled DNA sample on an array comprising a plurality of probes for said DNA under conditions which allow specific hybridization between said plurality of probes and said DNA; and (e)detecting said hybridization, wherein an amount of said label is indicative of the CpG methylation status in said DNA sample.

Provided herein are methods, compositions and kits for the generation of bisulfite-converted next generation sequencing (NGS) libraries. The methods, compositions and kits provided herein can be useful, for example, for the production of libraries from genomic DNA that allow for determination of the methylation status across the genome, i.e. the methylome. The methods, compositions and kits provided herein can also be utilized to query methylation status at a particular genomic locus or loci. Moreover, the methods provided herein can be employed for high-throughput sequencing of bisulfite-converted DNA while maintaining the directional (strandedness) information of the original nucleic acid sample.

Provided herein are methods, compositions and kits for the generation of bisulfite-converted next generation sequencing (NGS) libraries. The methods, compositions and kits provided herein can be useful, for example, for the production of libraries from genomic DNA that allow for determination of the methylation status across the genome, i.e. the methylome. The methods, compositions and kits provided herein can also be utilized to query methylation status at a particular genomic locus or loci. Moreover, the methods provided herein can be employed for high-throughput sequencing of bisulfite-converted DNA while maintaining the directional (strandedness) information of the original nucleic acid sample.

Provided herein are methods, compositions and kits for the generation of bisulfite-converted next generation sequencing (NGS) libraries. The methods, compositions and kits provided herein can be useful, for example, for the production of libraries from genomic DNA that allow for determination of the methylation status across the genome, i.e. the methylome. The methods, compositions and kits provided herein can also be utilized to query methylation status at a particular genomic locus or loci. Moreover, the methods provided herein can be employed for high-throughput sequencing of bisulfite-converted DNA while maintaining the directional (strandedness) information of the original nucleic acid sample.

The present invention provides assays and assay systems for use in spatially encoded biological assays. The invention provides an assay system comprising an assay capable of high levels of multiplexing where reagents are provided to a biological sample in defined spatial patterns; instrumentation capable of controlled delivery of reagents according to the spatial patterns; and a decoding scheme providing a readout that is digital in nature.

In one aspect, the invention features a combination of oligonucleotides comprising a forward primer oligonucleotide and a blocking oligonucleotide. The forward primer oligonucleotide has a 3′ end region, where the 3′ end region includes a portion complementary to a mutation positioned in a region within a polynucleotide. The blocking oligonucleotide contains a blocking moiety and has a 5′ end region, where the 5′ end region includes a portion complementary to a wild-type sequence of the region corresponding to the position of the mutation. In other aspects, the invention provides kits including the combination of primer oligonucleotides and methods of using the oligonucleotides to detect a mutation in a polynucleotide.

In one aspect, the invention features a combination of oligonucleotides comprising a forward primer oligonucleotide and a blocking oligonucleotide. The forward primer oligonucleotide has a 3′ end region, where the 3′ end region includes a portion complementary to a mutation positioned in a region within a polynucleotide. The blocking oligonucleotide contains a blocking moiety and has a 5′ end region, where the 5′ end region includes a portion complementary to a wild-type sequence of the region corresponding to the position of the mutation. In other aspects, the invention provides kits including the combination of primer oligonucleotides and methods of using the oligonucleotides to detect a mutation in a polynucleotide.

The present invention determines lymphocyte clonality by contacting a sample comprising nucleic acid molecules (1) of lymphocytes with forward and reverse primers (10, 20) and amplifying the nucleic acid molecules (1) by performing PCR pre-amplification to form barcoded PCR products (50). The barcoded PCR products (50) are amplified using adapter-specific forward and reverse primers (30, 40) in a PCR application into amplified barcoded PCR products (60), which are sequenced. The sequence reads are demultiplexed, mapped to respective TCR or BCR clonotypes and used to determine lymphocyte clonality for the sample. The forward and/or reverse primers (10, 20) are barcoded by comprising UMIs (14, 24) protected inside hairpin loops.

The present invention determines lymphocyte clonality by contacting a sample comprising nucleic acid molecules (1) of lymphocytes with forward and reverse primers (10, 20) and amplifying the nucleic acid molecules (1) by performing PCR pre-amplification to form barcoded PCR products (50). The barcoded PCR products (50) are amplified using adapter-specific forward and reverse primers (30, 40) in a PCR application into amplified barcoded PCR products (60), which are sequenced. The sequence reads are demultiplexed, mapped to respective TCR or BCR clonotypes and used to determine lymphocyte clonality for the sample. The forward and/or reverse primers (10, 20) are barcoded by comprising UMIs (14, 24) protected inside hairpin loops.

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.