Dataset Information

Heap: a highly sensitive and accurate SNP detection tool for low-coverage high-throughput sequencing data.

ABSTRACT: Recent availability of large-scale genomic resources enables us to conduct so called genome-wide association studies (GWAS) and genomic prediction (GP) studies, particularly with next-generation sequencing (NGS) data. The effectiveness of GWAS and GP depends on not only their mathematical models, but the quality and quantity of variants employed in the analysis. In NGS single nucleotide polymorphism (SNP) calling, conventional tools ideally require more reads for higher SNP sensitivity and accuracy. In this study, we aimed to develop a tool, Heap, that enables robustly sensitive and accurate calling of SNPs, particularly with a low coverage NGS data, which must be aligned to the reference genome sequences in advance. To reduce false positive SNPs, Heap determines genotypes and calls SNPs at each site except for sites at the both ends of reads or containing a minor allele supported by only one read. Performance comparison with existing tools showed that Heap achieved the highest F-scores with low coverage (7X) restriction-site associated DNA sequencing reads of sorghum and rice individuals. This will facilitate cost-effective GWAS and GP studies in this NGS era. Code and documentation of Heap are freely available from https://github.com/meiji-bioinf/heap (29 March 2017, date last accessed) and our web site (http://bioinf.mind.meiji.ac.jp/lab/en/tools.html (29 March 2017, date last accessed)).

SUBMITTER: Kobayashi M

PROVIDER: S-EPMC5737671 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature

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Heap: a highly sensitive and accurate SNP detection tool for low-coverage high-throughput sequencing data.

Kobayashi Masaaki M Ohyanagi Hajime H Takanashi Hideki H Asano Satomi S Kudo Toru T Kajiya-Kanegae Hiromi H Nagano Atsushi J AJ Tainaka Hitoshi H Tokunaga Tsuyoshi T Sazuka Takashi T Iwata Hiroyoshi H Tsutsumi Nobuhiro N Yano Kentaro K

DNA research : an international journal for rapid publication of reports on genes and genomes 20170801 4

Recent availability of large-scale genomic resources enables us to conduct so called genome-wide association studies (GWAS) and genomic prediction (GP) studies, particularly with next-generation sequencing (NGS) data. The effectiveness of GWAS and GP depends on not only their mathematical models, but the quality and quantity of variants employed in the analysis. In NGS single nucleotide polymorphism (SNP) calling, conventional tools ideally require more reads for higher SNP sensitivity and accur ...[more]

PMID: 28498906

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Project description:BACKGROUND: Many Single Nucleotide Polymorphism (SNP) calling programs have been developed to identify Single Nucleotide Variations (SNVs) in next-generation sequencing (NGS) data. However, low sequencing coverage presents challenges to accurate SNV identification, especially in single-sample data. Moreover, commonly used SNP calling programs usually include several metrics in their output files for each potential SNP. These metrics are highly correlated in complex patterns, making it extremely difficult to select SNPs for further experimental validations. RESULTS: To explore solutions to the above challenges, we compare the performance of four SNP calling algorithm, SOAPsnp, Atlas-SNP2, SAMtools, and GATK, in a low-coverage single-sample sequencing dataset. Without any post-output filtering, SOAPsnp calls more SNVs than the other programs since it has fewer internal filtering criteria. Atlas-SNP2 has stringent internal filtering criteria; thus it reports the least number of SNVs. The numbers of SNVs called by GATK and SAMtools fall between SOAPsnp and Atlas-SNP2. Moreover, we explore the values of key metrics related to SNVs' quality in each algorithm and use them as post-output filtering criteria to filter out low quality SNVs. Under different coverage cutoff values, we compare four algorithms and calculate the empirical positive calling rate and sensitivity. Our results show that: 1) the overall agreement of the four calling algorithms is low, especially in non-dbSNPs; 2) the agreement of the four algorithms is similar when using different coverage cutoffs, except that the non-dbSNPs agreement level tends to increase slightly with increasing coverage; 3) SOAPsnp, SAMtools, and GATK have a higher empirical calling rate for dbSNPs compared to non-dbSNPs; and 4) overall, GATK and Atlas-SNP2 have a relatively higher positive calling rate and sensitivity, but GATK calls more SNVs. CONCLUSIONS: Our results show that the agreement between different calling algorithms is relatively low. Thus, more caution should be used in choosing algorithms, setting filtering parameters, and designing validation studies. For reliable SNV calling results, we recommend that users employ more than one algorithm and use metrics related to calling quality and coverage as filtering criteria.

Project description:Targeted mass spectrometry (MS) approaches, which are powerful methods for uniquely and confidently quantifying a specific panel of proteins in complex biological samples, play a crucial role in validation and clinical translation of protein biomarkers discovered by global proteomic profiling. Common targeted MS methods, such as multiple reaction monitoring (MRM) and parallel-reaction monitoring (PRM), employ specific mass spectrometric technologies to quantify protein levels by comparing the transitions of specific endogenous (Endo) peptides with those of stable isotope labeled (SIL) peptide counterparts. These methods utilizing amino acid analyzed (AAA) SIL peptides warrants sensitive and precise measurements required for targeted MS assays. Compared to MRM, PRM provides higher experimental throughput by simultaneously acquiring all transitions of the target peptides and thereby compensate for different ion suppression among transitions of a target peptide. However, PRM still suffers different ion suppressions between Endo and SIL peptides due to poor spray stability as the Endo and SIL peptides were monitored at different liquid chromatography (LC) retention times. Here we introduce a new targeted MS method, termed as wideband PRM (WBPRM), designed for high-throughput targeted MS approach. WBPRM employs a wide isolation window for simultaneous fragmentations of both Endo and SIL peptides along with multiplexed single ion monitoring (SIM) scans for enhanced MS sensitivity of target peptides. Compared to PRM, WBPRM was demonstrated to provide increased sensitivity, precision, and accuracy of quantitative measurements of target peptides with increased throughput, allowing more target peptide measurements in a short experiment time. Its adaptability to a MS method provided by the manufacture makes it a facile method to implement for MS based assays, particularly in complex biological samples, where the needs for higher accuracy, sensitivity and efficiency are paramount.

Dataset Information

Heap: a highly sensitive and accurate SNP detection tool for low-coverage high-throughput sequencing data.

Publications

Heap: a highly sensitive and accurate SNP detection tool for low-coverage high-throughput sequencing data.

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