16,740 research outputs found
Discovery of large genomic inversions using long range information.
BackgroundAlthough many algorithms are now available that aim to characterize different classes of structural variation, discovery of balanced rearrangements such as inversions remains an open problem. This is mainly due to the fact that breakpoints of such events typically lie within segmental duplications or common repeats, which reduces the mappability of short reads. The algorithms developed within the 1000 Genomes Project to identify inversions are limited to relatively short inversions, and there are currently no available algorithms to discover large inversions using high throughput sequencing technologies.ResultsHere we propose a novel algorithm, VALOR, to discover large inversions using new sequencing methods that provide long range information such as 10X Genomics linked-read sequencing, pooled clone sequencing, or other similar technologies that we commonly refer to as long range sequencing. We demonstrate the utility of VALOR using both pooled clone sequencing and 10X Genomics linked-read sequencing generated from the genome of an individual from the HapMap project (NA12878). We also provide a comprehensive comparison of VALOR against several state-of-the-art structural variation discovery algorithms that use whole genome shotgun sequencing data.ConclusionsIn this paper, we show that VALOR is able to accurately discover all previously identified and experimentally validated large inversions in the same genome with a low false discovery rate. Using VALOR, we also predicted a novel inversion, which we validated using fluorescent in situ hybridization. VALOR is available at https://github.com/BilkentCompGen/VALOR
Jabba: hybrid error correction for long sequencing reads using maximal exact matches
Third generation sequencing platforms produce longer reads with higher error rates than second generation sequencing technologies. While the improved read length can provide useful information for downstream analysis, underlying algorithms are challenged by the high error rate. Error correction methods in which accurate short reads are used to correct noisy long reads appear to be attractive to generate high-quality long reads. Methods that align short reads to long reads do not optimally use the information contained in the second generation data, and suffer from large runtimes. Recently, a new hybrid error correcting method has been proposed, where the second generation data is first assembled into a de Bruijn graph, on which the long reads are then aligned. In this context we present Jabba, a hybrid method to correct long third generation reads by mapping them on a corrected de Bruijn graph that was constructed from second generation data. Unique to our method is that this mapping is constructed with a seed and extend methodology, using maximal exact matches as seeds. In addition to benchmark results, certain theoretical results concerning the possibilities and limitations of the use of maximal exact matches in the context of third generation reads are presented
Jabba: hybrid error correction for long sequencing reads
Background: Third generation sequencing platforms produce longer reads with higher error rates than second generation technologies. While the improved read length can provide useful information for downstream analysis, underlying algorithms are challenged by the high error rate. Error correction methods in which accurate short reads are used to correct noisy long reads appear to be attractive to generate high-quality long reads. Methods that align short reads to long reads do not optimally use the information contained in the second generation data, and suffer from large runtimes. Recently, a new hybrid error correcting method has been proposed, where the second generation data is first assembled into a de Bruijn graph, on which the long reads are then aligned.
Results: In this context we present Jabba, a hybrid method to correct long third generation reads by mapping them on a corrected de Bruijn graph that was constructed from second generation data. Unique to our method is the use of a pseudo alignment approach with a seed-and-extend methodology, using maximal exact matches (MEMs) as seeds. In addition to benchmark results, certain theoretical results concerning the possibilities and limitations of the use of MEMs in the context of third generation reads are presented.
Conclusion: Jabba produces highly reliable corrected reads: almost all corrected reads align to the reference, and these alignments have a very high identity. Many of the aligned reads are error-free. Additionally, Jabba corrects reads using a very low amount of CPU time. From this we conclude that pseudo alignment with MEMs is a fast and reliable method to map long highly erroneous sequences on a de Bruijn graph
A Reference-Free Algorithm for Computational Normalization of Shotgun Sequencing Data
Deep shotgun sequencing and analysis of genomes, transcriptomes, amplified
single-cell genomes, and metagenomes has enabled investigation of a wide range
of organisms and ecosystems. However, sampling variation in short-read data
sets and high sequencing error rates of modern sequencers present many new
computational challenges in data interpretation. These challenges have led to
the development of new classes of mapping tools and {\em de novo} assemblers.
These algorithms are challenged by the continued improvement in sequencing
throughput. We here describe digital normalization, a single-pass computational
algorithm that systematizes coverage in shotgun sequencing data sets, thereby
decreasing sampling variation, discarding redundant data, and removing the
majority of errors. Digital normalization substantially reduces the size of
shotgun data sets and decreases the memory and time requirements for {\em de
novo} sequence assembly, all without significantly impacting content of the
generated contigs. We apply digital normalization to the assembly of microbial
genomic data, amplified single-cell genomic data, and transcriptomic data. Our
implementation is freely available for use and modification
Nanopore Sequencing Technology and Tools for Genome Assembly: Computational Analysis of the Current State, Bottlenecks and Future Directions
Nanopore sequencing technology has the potential to render other sequencing
technologies obsolete with its ability to generate long reads and provide
portability. However, high error rates of the technology pose a challenge while
generating accurate genome assemblies. The tools used for nanopore sequence
analysis are of critical importance as they should overcome the high error
rates of the technology. Our goal in this work is to comprehensively analyze
current publicly available tools for nanopore sequence analysis to understand
their advantages, disadvantages, and performance bottlenecks. It is important
to understand where the current tools do not perform well to develop better
tools. To this end, we 1) analyze the multiple steps and the associated tools
in the genome assembly pipeline using nanopore sequence data, and 2) provide
guidelines for determining the appropriate tools for each step. We analyze
various combinations of different tools and expose the tradeoffs between
accuracy, performance, memory usage and scalability. We conclude that our
observations can guide researchers and practitioners in making conscious and
effective choices for each step of the genome assembly pipeline using nanopore
sequence data. Also, with the help of bottlenecks we have found, developers can
improve the current tools or build new ones that are both accurate and fast, in
order to overcome the high error rates of the nanopore sequencing technology.Comment: To appear in Briefings in Bioinformatics (BIB), 201
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