title: MetCap: a bioinformatics probe design pipeline for large-scale targeted metagenomics.
abstract: BACKGROUND
  Massive sequencing of genes from different environments has evolved metagenomics as central to enhancing the understanding of the wide diversity of micro-organisms and their roles in driving ecological processes. Reduced cost and high throughput sequencing has made large-scale projects achievable to a wider group of researchers, though complete metagenome sequencing is still a daunting task in terms of sequencing as well as the downstream bioinformatics analyses. Alternative approaches such as targeted amplicon sequencing requires custom PCR primer generation, and is not scalable to thousands of genes or gene families.
  
  
  RESULTS
  In this study, we are presenting a web-based tool called MetCap that circumvents the limitations of amplicon sequencing of multiple genes by designing probes that are suitable for large-scale targeted metagenomics sequencing studies. MetCap provides a novel approach to target thousands of genes and genomic regions that could be used in targeted metagenomics studies. Automatic analysis of user-defined sequences is performed, and probes specifically designed for metagenome studies are generated. To illustrate the advantage of a targeted metagenome approach, we have generated more than 400,000 probes that match more than 300,000 [corrected] publicly available sequences related to carbon degradation, and used these probes for target sequencing in a soil metagenome study. The results show high enrichment of target genes and a successful capturing of the majority of gene families. MetCap is freely available to users from: http://soilecology.biol.lu.se/metcap/ .
  
  
  CONCLUSIONS
  MetCap is facilitating probe-based target enrichment as an easy and efficient alternative tool compared to complex primer-based enrichment for large-scale investigations of metagenomes. Our results have shown efficient large-scale target enrichment through MetCap-designed probes for a soil metagenome. The web service is suitable for any targeted metagenomics project that aims to study several genes simultaneously. The novel bioinformatics approach taken by the web service will enable researchers in microbial ecology to tap into the vast diversity of microbial communities using targeted metagenomics as a cost-effective alternative to whole metagenome sequencing.
authors.affiliation: Department of Biology, Lund University, Ecology Building, 223 62, Lund, Sweden. sandeep.kushwaha@biol.lu.se.
authors.forename: Sandeep K
authors.initials: SK
authors.lastname: Kushwaha
authors.affiliation: Department of Biology, Lund University, Ecology Building, 223 62, Lund, Sweden. lokeshwaran.manoharan@biol.lu.se.
authors.forename: Lokeshwaran
authors.initials: L
authors.lastname: Manoharan
authors.affiliation: Department of Biology, Lund University, Ecology Building, 223 62, Lund, Sweden. tejashwari.meerupati@biol.lu.se.
authors.forename: Tejashwari
authors.initials: T
authors.lastname: Meerupati
authors.affiliation: Department of Biology, Lund University, Ecology Building, 223 62, Lund, Sweden. katarina.hedlund@biol.lu.se.
authors.forename: Katarina
authors.initials: K
authors.lastname: Hedlund
authors.affiliation: Department of Biology, Lund University, Ecology Building, 223 62, Lund, Sweden. dag.ahren@biol.lu.se.|Bioinformatics Infrastructure for Life Sciences (BILS), Department of Biology, Lund University, Ecology Building, 223 62, Lund, Sweden. dag.ahren@biol.lu.se.
authors.forename: Dag
authors.initials: D
authors.lastname: Ahrén
mesh_terms: D019295:Computational Biology / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004463:Ecology; D004777:Environment; D054892:Metagenome; D056186:Metagenomics / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods; D012984:Software; D012987:Soil / Q000737:chemistry*
chemical_list: D017931:DNA Primers; D012987:Soil