3,539 research outputs found
SOAP3-dp: Fast, Accurate and Sensitive GPU-based Short Read Aligner
To tackle the exponentially increasing throughput of Next-Generation
Sequencing (NGS), most of the existing short-read aligners can be configured to
favor speed in trade of accuracy and sensitivity. SOAP3-dp, through leveraging
the computational power of both CPU and GPU with optimized algorithms, delivers
high speed and sensitivity simultaneously. Compared with widely adopted
aligners including BWA, Bowtie2, SeqAlto, GEM and GPU-based aligners including
BarraCUDA and CUSHAW, SOAP3-dp is two to tens of times faster, while
maintaining the highest sensitivity and lowest false discovery rate (FDR) on
Illumina reads with different lengths. Transcending its predecessor SOAP3,
which does not allow gapped alignment, SOAP3-dp by default tolerates alignment
similarity as low as 60 percent. Real data evaluation using human genome
demonstrates SOAP3-dp's power to enable more authentic variants and longer
Indels to be discovered. Fosmid sequencing shows a 9.1 percent FDR on newly
discovered deletions. SOAP3-dp natively supports BAM file format and provides a
scoring scheme same as BWA, which enables it to be integrated into existing
analysis pipelines. SOAP3-dp has been deployed on Amazon-EC2, NIH-Biowulf and
Tianhe-1A.Comment: 21 pages, 6 figures, submitted to PLoS ONE, additional files
available at "https://www.dropbox.com/sh/bhclhxpoiubh371/O5CO_CkXQE".
Comments most welcom
MaxSSmap: A GPU program for mapping divergent short reads to genomes with the maximum scoring subsequence
Programs based on hash tables and Burrows-Wheeler are very fast for mapping
short reads to genomes but have low accuracy in the presence of mismatches and
gaps. Such reads can be aligned accurately with the Smith-Waterman algorithm
but it can take hours and days to map millions of reads even for bacteria
genomes. We introduce a GPU program called MaxSSmap with the aim of achieving
comparable accuracy to Smith-Waterman but with faster runtimes. Similar to most
programs MaxSSmap identifies a local region of the genome followed by exact
alignment. Instead of using hash tables or Burrows-Wheeler in the first part,
MaxSSmap calculates maximum scoring subsequence score between the read and
disjoint fragments of the genome in parallel on a GPU and selects the highest
scoring fragment for exact alignment. We evaluate MaxSSmap's accuracy and
runtime when mapping simulated Illumina E.coli and human chromosome one reads
of different lengths and 10\% to 30\% mismatches with gaps to the E.coli genome
and human chromosome one. We also demonstrate applications on real data by
mapping ancient horse DNA reads to modern genomes and unmapped paired reads
from NA12878 in 1000 genomes. We show that MaxSSmap attains comparable high
accuracy and low error to fast Smith-Waterman programs yet has much lower
runtimes. We show that MaxSSmap can map reads rejected by BWA and NextGenMap
with high accuracy and low error much faster than if Smith-Waterman were used.
On short read lengths of 36 and 51 both MaxSSmap and Smith-Waterman have lower
accuracy compared to at higher lengths. On real data MaxSSmap produces many
alignments with high score and mapping quality that are not given by NextGenMap
and BWA. The MaxSSmap source code is freely available from
http://www.cs.njit.edu/usman/MaxSSmap
Technology dictates algorithms: Recent developments in read alignment
Massively parallel sequencing techniques have revolutionized biological and
medical sciences by providing unprecedented insight into the genomes of humans,
animals, and microbes. Modern sequencing platforms generate enormous amounts of
genomic data in the form of nucleotide sequences or reads. Aligning reads onto
reference genomes enables the identification of individual-specific genetic
variants and is an essential step of the majority of genomic analysis
pipelines. Aligned reads are essential for answering important biological
questions, such as detecting mutations driving various human diseases and
complex traits as well as identifying species present in metagenomic samples.
The read alignment problem is extremely challenging due to the large size of
analyzed datasets and numerous technological limitations of sequencing
platforms, and researchers have developed novel bioinformatics algorithms to
tackle these difficulties. Importantly, computational algorithms have evolved
and diversified in accordance with technological advances, leading to todays
diverse array of bioinformatics tools. Our review provides a survey of
algorithmic foundations and methodologies across 107 alignment methods
published between 1988 and 2020, for both short and long reads. We provide
rigorous experimental evaluation of 11 read aligners to demonstrate the effect
of these underlying algorithms on speed and efficiency of read aligners. We
separately discuss how longer read lengths produce unique advantages and
limitations to read alignment techniques. We also discuss how general alignment
algorithms have been tailored to the specific needs of various domains in
biology, including whole transcriptome, adaptive immune repertoire, and human
microbiome studies
SOAP3-dp: Fast, Accurate and Sensitive GPU-Based Short Read Aligner
To tackle the exponentially increasing throughput of Next-Generation Sequencing (NGS), most of the existing short-read aligners can be configured to favor speed in trade of accuracy and sensitivity. SOAP3-dp, through leveraging the computational power of both CPU and GPU with optimized algorithms, delivers high speed and sensitivity simultaneously. Compared with widely adopted aligners including BWA, Bowtie2, SeqAlto, CUSHAW2, GEM and GPU-based aligners BarraCUDA and CUSHAW, SOAP3-dp was found to be two to tens of times faster, while maintaining the highest sensitivity and lowest false discovery rate (FDR) on Illumina reads with different lengths. Transcending its predecessor SOAP3, which does not allow gapped alignment, SOAP3-dp by default tolerates alignment similarity as low as 60%. Real data evaluation using human genome demonstrates SOAP3-dp's power to enable more authentic variants and longer Indels to be discovered. Fosmid sequencing shows a 9.1% FDR on newly discovered deletions. SOAP3-dp natively supports BAM file format and provides the same scoring scheme as BWA, which enables it to be integrated into existing analysis pipelines. SOAP3-dp has been deployed on Amazon-EC2, NIH-Biowulf and Tianhe-1A
Virtual Environment for Next Generation Sequencing Analysis
Next Generation Sequencing technology, on the one hand, allows a more accurate analysis, and, on the other hand, increases the amount of data to process. A new protocol for sequencing the messenger RNA in a cell, known as RNA- Seq, generates millions of short sequence fragments in a single run. These fragments, or reads, can be used to measure levels of gene expression and to identify novel splice variants of genes. The proposed solution is a distributed architecture consisting of a Grid Environment and a Virtual Grid Environment, in order to reduce processing time by making the system scalable and flexibl
Extreme Scale De Novo Metagenome Assembly
Metagenome assembly is the process of transforming a set of short,
overlapping, and potentially erroneous DNA segments from environmental samples
into the accurate representation of the underlying microbiomes's genomes.
State-of-the-art tools require big shared memory machines and cannot handle
contemporary metagenome datasets that exceed Terabytes in size. In this paper,
we introduce the MetaHipMer pipeline, a high-quality and high-performance
metagenome assembler that employs an iterative de Bruijn graph approach.
MetaHipMer leverages a specialized scaffolding algorithm that produces long
scaffolds and accommodates the idiosyncrasies of metagenomes. MetaHipMer is
end-to-end parallelized using the Unified Parallel C language and therefore can
run seamlessly on shared and distributed-memory systems. Experimental results
show that MetaHipMer matches or outperforms the state-of-the-art tools in terms
of accuracy. Moreover, MetaHipMer scales efficiently to large concurrencies and
is able to assemble previously intractable grand challenge metagenomes. We
demonstrate the unprecedented capability of MetaHipMer by computing the first
full assembly of the Twitchell Wetlands dataset, consisting of 7.5 billion
reads - size 2.6 TBytes.Comment: Accepted to SC1
An improved genome of the model marine alga Ostreococcus tauri unfolds by assessing Illumina de novo assemblies
Background:
Cost effective next generation sequencing technologies now enable the production of genomic datasets for many novel planktonic eukaryotes, representing an understudied reservoir of genetic diversity. O. tauri is the smallest free-living photosynthetic eukaryote known to date, a coccoid green alga that was first isolated in 1995 in a lagoon by the Mediterranean sea. Its simple features, ease of culture and the sequencing of its 13 Mb haploid nuclear genome have promoted this microalga as a new model organism for cell biology. Here, we investigated the quality of genome assemblies of Illumina GAIIx 75 bp paired-end reads from Ostreococcus tauri, thereby also improving the existing assembly and showing the genome to be stably maintained in culture.
Results:
The 3 assemblers used, ABySS, CLCBio and Velvet, produced 95% complete genomes in 1402 to 2080 scaffolds with a very low rate of misassembly. Reciprocally, these assemblies improved the original genome assembly by filling in 930 gaps. Combined with additional analysis of raw reads and PCR sequencing effort, 1194 gaps have been solved in total adding up to 460 kb of sequence. Mapping of RNAseq Illumina data on this updated genome led to a twofold reduction in the proportion of multi-exon protein coding genes, representing 19% of the total 7699 protein coding genes. The comparison of the DNA extracted in 2001 and 2009 revealed the fixation of 8 single nucleotide substitutions and 2 deletions during the approximately 6000 generations in the lab. The deletions either knocked out or truncated two predicted transmembrane proteins, including a glutamate-receptor like gene.
Conclusion:
High coverage (>80 fold) paired-end Illumina sequencing enables a high quality 95% complete genome assembly of a compact ~13 Mb haploid eukaryote. This genome sequence has remained stable for 6000 generations of lab culture
NOVEL COMPUTATIONAL METHODS FOR SEQUENCING DATA ANALYSIS: MAPPING, QUERY, AND CLASSIFICATION
Over the past decade, the evolution of next-generation sequencing technology has considerably advanced the genomics research. As a consequence, fast and accurate computational methods are needed for analyzing the large data in different applications. The research presented in this dissertation focuses on three areas: RNA-seq read mapping, large-scale data query, and metagenomics sequence classification.
A critical step of RNA-seq data analysis is to map the RNA-seq reads onto a reference genome. This dissertation presents a novel splice alignment tool, MapSplice3. It achieves high read alignment and base mapping yields and is able to detect splice junctions, gene fusions, and circular RNAs comprehensively at the same time. Based on MapSplice3, we further extend a novel lightweight approach called iMapSplice that enables personalized mRNA transcriptional profiling. As huge amount of RNA-seq has been shared through public datasets, it provides invaluable resources for researchers to test hypotheses by reusing existing datasets. To meet the needs of efficiently querying large-scale sequencing data, a novel method, called SeqOthello, has been developed. It is able to efficiently query sequence k-mers against large-scale datasets and finally determines the existence of the given sequence. Metagenomics studies often generate tens of millions of reads to capture the presence of microbial organisms. Thus efficient and accurate algorithms are in high demand. In this dissertation, we introduce MetaOthello, a probabilistic hashing classifier for metagenomic sequences. It supports efficient query of a taxon using its k-mer signatures
From cheek swabs to consensus sequences : an A to Z protocol for high-throughput DNA sequencing of complete human mitochondrial genomes
Background: Next-generation DNA sequencing (NGS) technologies have made huge impacts in many fields of biological research, but especially in evolutionary biology. One area where NGS has shown potential is for high-throughput sequencing of complete mtDNA genomes (of humans and other animals). Despite the increasing use of NGS technologies and a better appreciation of their importance in answering biological questions, there remain significant obstacles to the successful implementation of NGS-based projects, especially for new users.
Results: Here we present an ‘A to Z’ protocol for obtaining complete human mitochondrial (mtDNA) genomes – from DNA extraction to consensus sequence. Although designed for use on humans, this protocol could also be used to sequence small, organellar genomes from other species, and also nuclear loci. This protocol includes DNA extraction, PCR amplification, fragmentation of PCR products, barcoding of fragments, sequencing using the 454 GS FLX platform, and a complete bioinformatics pipeline (primer removal, reference-based mapping, output of coverage plots and SNP calling).
Conclusions: All steps in this protocol are designed to be straightforward to implement, especially for researchers who are undertaking next-generation sequencing for the first time. The molecular steps are scalable to large numbers (hundreds) of individuals and all steps post-DNA extraction can be carried out in 96-well plate format. Also, the protocol has been assembled so that individual ‘modules’ can be swapped out to suit available resources
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