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An improved filtering algorithm for big read datasets and its application to single-cell assembly

BACKGROUND: For single-cell or metagenomic sequencing projects, it is necessary to sequence with a very high mean coverage in order to make sure that all parts of the sample DNA get covered by the reads produced. This leads to huge datasets with lots of redundant data. A filtering of this data prior...

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Detalles Bibliográficos
Autores principales: Wedemeyer, Axel, Kliemann, Lasse, Srivastav, Anand, Schielke, Christian, Reusch, Thorsten B., Rosenstiel, Philip
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5496428/
https://www.ncbi.nlm.nih.gov/pubmed/28673253
http://dx.doi.org/10.1186/s12859-017-1724-7
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author Wedemeyer, Axel
Kliemann, Lasse
Srivastav, Anand
Schielke, Christian
Reusch, Thorsten B.
Rosenstiel, Philip
author_facet Wedemeyer, Axel
Kliemann, Lasse
Srivastav, Anand
Schielke, Christian
Reusch, Thorsten B.
Rosenstiel, Philip
author_sort Wedemeyer, Axel
collection PubMed
description BACKGROUND: For single-cell or metagenomic sequencing projects, it is necessary to sequence with a very high mean coverage in order to make sure that all parts of the sample DNA get covered by the reads produced. This leads to huge datasets with lots of redundant data. A filtering of this data prior to assembly is advisable. Brown et al. (2012) presented the algorithm Diginorm for this purpose, which filters reads based on the abundance of their k-mers. METHODS: We present Bignorm, a faster and quality-conscious read filtering algorithm. An important new algorithmic feature is the use of phred quality scores together with a detailed analysis of the k-mer counts to decide which reads to keep. RESULTS: We qualify and recommend parameters for our new read filtering algorithm. Guided by these parameters, we remove in terms of median 97.15% of the reads while keeping the mean phred score of the filtered dataset high. Using the SDAdes assembler, we produce assemblies of high quality from these filtered datasets in a fraction of the time needed for an assembly from the datasets filtered with Diginorm. CONCLUSIONS: We conclude that read filtering is a practical and efficient method for reducing read data and for speeding up the assembly process. This applies not only for single cell assembly, as shown in this paper, but also to other projects with high mean coverage datasets like metagenomic sequencing projects. Our Bignorm algorithm allows assemblies of competitive quality in comparison to Diginorm, while being much faster. Bignorm is available for download at https://git.informatik.uni-kiel.de/axw/Bignorm. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12859-017-1724-7) contains supplementary material, which is available to authorized users.
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spelling pubmed-54964282017-07-07 An improved filtering algorithm for big read datasets and its application to single-cell assembly Wedemeyer, Axel Kliemann, Lasse Srivastav, Anand Schielke, Christian Reusch, Thorsten B. Rosenstiel, Philip BMC Bioinformatics Methodology Article BACKGROUND: For single-cell or metagenomic sequencing projects, it is necessary to sequence with a very high mean coverage in order to make sure that all parts of the sample DNA get covered by the reads produced. This leads to huge datasets with lots of redundant data. A filtering of this data prior to assembly is advisable. Brown et al. (2012) presented the algorithm Diginorm for this purpose, which filters reads based on the abundance of their k-mers. METHODS: We present Bignorm, a faster and quality-conscious read filtering algorithm. An important new algorithmic feature is the use of phred quality scores together with a detailed analysis of the k-mer counts to decide which reads to keep. RESULTS: We qualify and recommend parameters for our new read filtering algorithm. Guided by these parameters, we remove in terms of median 97.15% of the reads while keeping the mean phred score of the filtered dataset high. Using the SDAdes assembler, we produce assemblies of high quality from these filtered datasets in a fraction of the time needed for an assembly from the datasets filtered with Diginorm. CONCLUSIONS: We conclude that read filtering is a practical and efficient method for reducing read data and for speeding up the assembly process. This applies not only for single cell assembly, as shown in this paper, but also to other projects with high mean coverage datasets like metagenomic sequencing projects. Our Bignorm algorithm allows assemblies of competitive quality in comparison to Diginorm, while being much faster. Bignorm is available for download at https://git.informatik.uni-kiel.de/axw/Bignorm. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12859-017-1724-7) contains supplementary material, which is available to authorized users. BioMed Central 2017-07-03 /pmc/articles/PMC5496428/ /pubmed/28673253 http://dx.doi.org/10.1186/s12859-017-1724-7 Text en © The Author(s) 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License(http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology Article
Wedemeyer, Axel
Kliemann, Lasse
Srivastav, Anand
Schielke, Christian
Reusch, Thorsten B.
Rosenstiel, Philip
An improved filtering algorithm for big read datasets and its application to single-cell assembly
title An improved filtering algorithm for big read datasets and its application to single-cell assembly
title_full An improved filtering algorithm for big read datasets and its application to single-cell assembly
title_fullStr An improved filtering algorithm for big read datasets and its application to single-cell assembly
title_full_unstemmed An improved filtering algorithm for big read datasets and its application to single-cell assembly
title_short An improved filtering algorithm for big read datasets and its application to single-cell assembly
title_sort improved filtering algorithm for big read datasets and its application to single-cell assembly
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5496428/
https://www.ncbi.nlm.nih.gov/pubmed/28673253
http://dx.doi.org/10.1186/s12859-017-1724-7
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