Several new genomics technologies have become available that offer long-read sequencing or long-range mapping with higher throughput and higher resolution analysis than ever before. These long-range technologies are rapidly advancing the field with improved reference genomes, more comprehensive variant identification and more complete views of transcriptomes and epigenomes. However, they also require new bioinformatics approaches to take full advantage of their unique characteristics while overcoming their complex errors and modalities. Here, we discuss several of the most important applications of the new technologies, focusing on both the currently available bioinformatics tools and opportunities for future research.
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The authors thank A. Phillippy, W. Timp, W. R. McCombie, S. Goodwin and R. Gibbs for helpful discussions. This work was supported, in part, by awards from the National Science Foundation (DBI-1350041) and from the National Institutes of Health (R01-HG006677 and UM1-HG008898). Also, this work was completed in part while H.L. was visiting the Simons Institute for the Theory of Computing, University of California, Berkeley, USA.
M.C.S. and F.J.S. have participated in Pacific Biosciences (PacBio) sponsored meetings over the past few years and have received travel reimbursement and honoraria for presenting at these events. PacBio had no role in decisions relating to the study and/or work to be published, data collection and analysis or the decision to publish.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Nature Reviews thanks Heng Li, René Warren and the other anonymous reviewer(s) for their contribution to the peer review of this work.
10X Genomics: https://www.10xgenomics.com/
Bionano Genomics: https://bionanogenomics.com/
Oxford Nanopore Technologies: https://nanoporetech.com/
Pacific Biosciences: http://www.pacb.com/
Proposed solutions to corruption of long-read sequencing files: https://github.com/samtools/hts-specs/issues/40
- Mate pairs
A molecular technique to generate a pair of sequencing reads separated by an approximately known distance. The typical separation distance for mate pairs is a few kilobases, as opposed to paired-end sequencing, which separates the reads by a few hundred bases at most.
- Optical mapping
A microscopy technique used to visualize the characteristics of DNA, especially the physical lengths or the position of fluorescent probes.
A type of DNA sequence variation marked by the insertion or deletion of nucleotides.
- Structural variants
(SVs). DNA sequence variants that are 50 bp or larger, including insertions, deletions, inversions, duplications and translocations.
- Linked reads
Also known as a read cloud. A set of barcoded short reads derived from the same DNA molecule and therefore highly localized in the genome.
Grouping together variants located on the same molecule, such as to identify variants from the maternal or the paternal genome in a diploid sample.
The process of assembling sequences of DNA into a scaffold. A scaffold is similar to a contig but may contain gaps, typically represented as Ns in the sequence.
Contiguously assembled sequences of DNA.
A weighted average length; specifically, the N50 length is the length such that 50% of the genome has been assembled into contig or scaffold sequences of this length or longer.
Any molecular interaction that regulates the transcription of nearby genes on the same DNA molecule, such as the role of a gene promoter.
Any molecular interaction that regulates the transcription of genes on a different DNA molecule, such as a transcription factor regulating both alleles of a target gene or genes.
- Topologically associating domains
(TADs). Regions of the genome that are enriched for interactions with other elements within the same domain.
- Synteny blocks
Genomic regions that are conserved among multiple species.
- Fragile sites
Regions of the DNA molecule that are prone to physical shearing, especially when multiple nicking sites targeted by a nicking enzyme are located in close proximity.
Two or more adjacent or even overlapping variants in the same region of the genome, such as a deletion within the middle of a larger inverted sequence.
A phenomenon by which many chromosomal rearrangements occur in a single event in a localized region of the genome. Also called chromosome shattering.
A complex mutation where genetic material from multiple chromosomes is broken and ligated to each other in a new configuration, especially in cancer.
Cells and organisms that contain more than two paired (homologous) sets of chromosomes.
Variants observed in the genome that are present to some appreciable degree within a population (for example, >1%).
- Compound heterozygous
Two different mutant alleles at a particular gene locus, one on each chromosome of a pair.
- Hemizygous mutations
Two or more heterozygous mutations, especially loss-of-function mutations, occurring on the same chromosome so that they disrupt one copy of the gene but leave one functional copy.
- Private mutations
Rare variants observed only within a single person or family.
In computational complexity theory, this is a class of problems in which no fast solutions are known to exist.
The genomes of all the species present in a sample, studied without culturing or otherwise isolating any individual.
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Sedlazeck, F.J., Lee, H., Darby, C.A. et al. Piercing the dark matter: bioinformatics of long-range sequencing and mapping. Nat Rev Genet 19, 329–346 (2018). https://doi.org/10.1038/s41576-018-0003-4
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