Abstract: Provided are methods of nucleic acid sequencing. The methods include producing a circularized DNA including a full-length cDNA and a known heterologous sequence, and performing rolling circle amplification using the circularized DNA as template to produce a concatemer including repeating segments including the full-length cDNA and the known heterologous sequence. The methods further include obtaining a raw sequencing read of the concatemer using a nanopore, identifying the repeating segments in the raw sequencing read, and producing a consensus sequence of the full-length cDNA based on the sequences of the repeating segments. Computer-readable media, computing devices, and systems that find use, e.g., in practicing the methods of the present disclosure are also provided.

Abstract: Aspects of the present disclosure include methods of making barcoded solid supports. In some embodiments, the methods include producing a concatemer by rolling circle amplification (RCA) of a circular nucleic acid template, where the circular nucleic acid template includes a barcode and a stem-loop forming region, and where the concatemer includes a plurality of linked units, each unit including the barcode and a stem-loop structure formed from the stem-loop forming region. Such methods further include disposing the concatemer on a solid support to produce a barcoded solid support including a plurality of the stem-loop structures extending from the surface of the solid support. The methods may further include treating the stem-loop structures with an agent that produces stem structures having ends compatible with target nucleic acids, and attaching the target nucleic acids to the stem structures. Barcoded solid supports and methods of using the barcoded solid supports are also provided.

Abstract: Provided are computer-implemented methods for comparing genotype data from a first sample to a limited amount of DNA sequence data from a second sample. In certain embodiments, the first sample is from a known individual and the second sample is an unknown sample. The methods find use in a variety of contexts, including for genetic identity detection, e.g., for forensic and other applications. Also provided are computer-implemented methods for assessing the degree of relatedness between genotype data from a first sample and a limited amount of DNA sequence data from a second sample. Computer-readable media and systems that find use in practicing the methods of the present disclosure are also provided.

Abstract: Provided are methods of producing nucleic acid libraries. In certain aspects, the methods include combining target nucleic acids (e.g., 5? phosphorylated nucleic acids) and an oligonucleotide pool. Oligonucleotides of the oligonucleotide pool may include complementarity regions of varying length and nucleotide sequence, and a complementarity region identification sequence. In such aspects, the combining is under conditions in which oligonucleotides of the oligonucleotide pool hybridize to nucleic acids of the target nucleic acids (e.g., 5? phosphorylated nucleic acids) having overhang regions that are complementary in sequence and have corresponding lengths with respect to the complementarity regions of the oligonucleotides. Compositions and kits that find use, e.g., in practicing the methods of the present disclosure are also provided.

Abstract: Disclosed are methods for testing biological samples containing genomic nucleic acids obtained from an organism having a genome, such as a human genome. It is often desirable to analyze a DNA sample or more than one, different DNA samples, to determine whether the sample comes from one individual or two individuals. The present method requires very low amounts of DNA and can use partial sequences of DNA fragments. Partial sequences are analyzed for the presence of polymorphisms (e.g. SNP's) that can be mapped to a reference SNP map. The distance between similar SNPS, which are genetically linked, can be used to statistically determine a likelihood of identity of individuality in a sample.

Abstract: Disclosed are methods for testing biological samples containing genomic nucleic acids obtained from an organism having a genome, such as a human genome. It is often desirable to analyze a DNA sample or more than one, different DNA samples, to determine whether the sample comes from one individual or two individuals. The present method requires very low amounts of DNA and can use partial sequences of DNA fragments. Partial sequences are analyzed for the presence of polymorphisms (e.g. SNP's) that can be mapped to a reference SNP map. The distance between similar SNPS, which are genetically linked, can be used to statistically determine a likelihood of identity of individuality in a sample.

Abstract: Provided are methods of producing nucleic acid libraries. In certain aspects, the methods include combining target nucleic acids (e.g., 5? phosphorylated nucleic acids) and an oligonucleotide pool. Oligonucleotides of the oligonucleotide pool may include complementarity regions of varying length and nucleotide sequence, and a complementarity region identification sequence. In such aspects, the combining is under conditions in which oligonucleotides of the oligonucleotide pool hybridize to nucleic acids of the target nucleic acids (e.g., 5? phosphorylated nucleic acids) having overhang regions that are complementary in sequence and have corresponding lengths with respect to the complementarity regions of the oligonucleotides. Compositions and kits that find use, e.g., in practicing the methods of the present disclosure are also provided.

Abstract: Methods and compositions for the de novo generation of scaffold information, linkage information and genome information for unknown organisms in heterogeneous metagenomic samples or samples obtained from multiple individuals are disclosed. Methods of the disclosure use a combination of restriction enzymes that have different sensitivities to specific base modifications to generate Chicago libraries. Practice of the methods allows de novo sequencing of entire genomes of uncultured or unidentified organisms in heterogeneous samples, or the determination of linkage information for nucleic acid molecules in samples comprising nucleic acids obtained from multiple individuals.

Abstract: Provided are methods of nucleic acid sequencing. The methods include producing a circularized DNA including a full-length cDNA and a known heterologous sequence, and performing rolling circle amplification using the circularized DNA as template to produce a concatemer including repeating segments including the full-length cDNA and the known heterologous sequence. The methods further include obtaining a raw sequencing read of the concatemer using a nanopore, identifying the repeating segments in the raw sequencing read, and producing a consensus sequence of the full-length cDNA based on the sequences of the repeating segments. Computer-readable media, computing devices, and systems that find use, e.g., in practicing the methods of the present disclosure are also provided.

Abstract: Provided are methods of producing nucleic acid libraries. In certain aspects, the methods include combining target nucleic acids (e.g., 5? phosphorylated nucleic acids) and an oligonucleotide pool. Oligonucleotides of the oligonucleotide pool may include complementarity regions of varying length and nucleotide sequence, and a complementarity region identification sequence. In such aspects, the combining is under conditions in which oligonucleotides of the oligonucleotide pool hybridize to nucleic acids of the target nucleic acids (e.g., 5? phosphorylated nucleic acids) having overhang regions that are complementary in sequence and have corresponding lengths with respect to the complementarity regions of the oligonucleotides. Compositions and kits that find use, e.g., in practicing the methods of the present disclosure are also provided.

Abstract: The disclosure provides methods for haplotype phasing and meta-genomics assemblies. The disclosure provides a streamlined method for accomplishing these tasks, such that intermediates need not be labeled by an affinity label to facilitate binding to a solid surface. The disclosure also provides methods and compositions for the de novo generation of scaffold information, linkage information, and genome information for unknown organisms in heterogeneous metagenomic samples or samples obtained from multiple individuals. Practice of the methods can allow de novo sequencing of entire genomes of uncultured or unidentified organisms in heterogeneous samples, or the determination of linkage information for nucleic acid molecules in samples comprising nucleic acids obtained from multiple individuals.

Abstract: The disclosure provides methods to assemble genomes of eukaryotic or prokaryotic organisms. The disclosure provides methods for haplotype phasing and meta-genomics assemblies. The disclosure provides a streamlined method for accomplishing these tasks, such that intermediates need not be labeled by an affinity label to facilitate binding to a solid surface. The disclosure also provides methods and compositions for the de novo generation of scaffold information, linkage information, and genome information for unknown organisms in heterogeneous metagenomic samples or samples obtained from multiple individuals. Practice of the methods can allow de novo sequencing of entire genomes of uncultured or unidentified organisms in heterogeneous samples, or the determination of linkage information for nucleic acid molecules in samples comprising nucleic acids obtained from multiple individuals.

Abstract: The disclosure provides methods to assemble genomes of eukaryotic or prokaryotic organisms. The disclosure provides methods for haplotype phasing and meta-genomics assemblies. The disclosure provides a streamlined method for accomplishing these tasks, such that intermediates need not be labeled by an affinity label to facilitate binding to a solid surface. The disclosure also provides methods and compositions for the de novo generation of scaffold information, linkage information, and genome information for unknown organisms in heterogeneous metagenomic samples or samples obtained from multiple individuals. Practice of the methods can allow de novo sequencing of entire genomes of uncultured or unidentified organisms in heterogeneous samples, or the determination of linkage information for nucleic acid molecules in samples comprising nucleic acids obtained from multiple individuals.

Abstract: Disclosed are methods for testing biological samples containing genomic nucleic acids obtained from an organism having a genome, such as a human genome. It is often desirable to analyze a DNA sample or more than one, different DNA samples, to determine whether the sample comes from one individual or two individuals. The present method requires very low amounts of DNA and can use partial sequences of DNA fragments. Partial sequences are analyzed for the presence of polymorphisms (e.g. SNP's) that can be mapped to a reference SNP map. The distance between similar SNPS, which are genetically linked, can be used to statistically determine a likelihood of identity of individuality in a sample.

Abstract: The disclosure provides methods to assemble genomes of eukaryotic or prokaryotic organisms. The disclosure further provides methods for haplotype phasing and meta-genomics assemblies.

Abstract: A modification to a robot end of arm tool to improve the retention of pail goods during transport by the tool includes affixing to the lower surface of a of rigid, planar, substrate, one or more pail lifting apparatus, each apparatus including a plurality of pail engaging fingers and a spacer block mounted to the substrate between the plurality of fingers. Each finger includes a flipper arm that is selectively, hydraulically controllable to move from a vertical to a horizontal position. When the flipper arm is in the horizontal position, it is able to physically engage the outer surface or ledge of an associated pail to provide support to the pail. The spacer block, located above the lid of the associated pail, when the pail is engaged by the plurality of fingers, restricts vertical movement of the pail during transport. The combination of fingers and a spacer block form one pail lifting apparatus. Multiple such apparatus may be affixed to the tool substrate.

Abstract: Computational methods for systematically characterizing putative protein isoforms as apparent targets of nonsense-mediated decay (NMD) comprise: (a) identifying a dataset of target putative protein isoform sequences for characterization; (b) identifying from an mRNA dataset corresponding mRNA sequences representing transcripts encoding the protein isoforms; (c) determining corresponding gene intron-exon structures by mapping the mRNA sequences to corresponding genomic sequences; and (d) determining if the transcripts are apparent targets of NMD. Methods for regulating the expression of a gene encoding a protein isoform characterized as an apparent target of NMD comprise biasing expression of the isoform by modulating transcript splicing or modulating NMD activity.

Abstract: Computational methods identify alternate splice forms of known gene transcripts and isoforms that are subject to NMD (nonsense-mediated decay). These methods were used to identify thousands of human genes that generate alternative splice forms, and to demonstrate that about a third of these are subject to NMD. This high prevalence of NMD-targeted transcripts indicates a systemic way of regulating gene expression—by shunting gene expression to nonproductive splice variants. This endemic regulation is exploited to engineer regulation of gene expression, to characterize splice pathway components and to assay splice environments, for example, using NMD-regulated reporter genes.

Abstract: Computational methods identify alternate splice forms of known gene transcripts and isoforms that are subject to NMD (nonsense-mediated decay). These methods were used to identify thousands of human genes that generate alternative splice forms, and to demonstrate that about a third of these are subject to NMD. This high prevalence of NMD-targeted transcripts indicates a systemic way of regulating gene expression—by shunting gene expression to nonproductive splice variants. This endemic regulation is exploited to engineer regulation of gene expression, to characterize splice pathway components and to assay splice environments, for example, using NMD-regulated reporter genes.