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Curr Protoc Microbiol. 12 2 (2006).","journal-title":"Curr Protoc Microbiol"},{"issue":"10","key":"ref27","doi-asserted-by":"crossref","first-page":"1451","DOI":"10.1101\/gr.4086505","type":"journal-article","article-title":"Galaxy: a platform for interactive large-scale genome analysis","volume":"15","author":"Giardine","year":"2005","unstructured":"Giardine, B. et al. Galaxy: a platform for interactive large-scale genome analysis. Genome Res. 15 (10), 1451-1455 (2005).","journal-title":"Genome Res"},{"issue":"8","key":"ref28","doi-asserted-by":"crossref","first-page":"R86","DOI":"10.1186\/gb-2010-11-8-r86","type":"journal-article","article-title":"Galaxy: a comprehensive approach for supporting accessible, reproducible, and transparent computational research in the life sciences","volume":"11","author":"Goecks","year":"2010","unstructured":"Goecks, J., Nekrutenko, A., Taylor, J., & Galaxy, T. 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Nucleic Acids Res. 8 (44) (W1):W3-W10 (2016).","journal-title":"Nucleic Acids Res"},{"key":"ref31","first-page":"1367","type":"journal-article","article-title":"MAGenTA; a Galaxy implemented tool for complete Tn-Seq analysis and data visualization","author":"McCoy","year":"2017","unstructured":"McCoy, K. M., Antonio, M. L., & van Opijnen, T. MAGenTA; a Galaxy implemented tool for complete Tn-Seq analysis and data visualization. Bioinformatics. 1367-4811 (Electronic) (2017).","journal-title":"Bioinformatics"},{"issue":"3","key":"ref32","doi-asserted-by":"crossref","first-page":"R25","DOI":"10.1186\/gb-2009-10-3-r25","type":"journal-article","article-title":"Ultrafast and memory-efficient alignment of short DNA sequences to the human genome","volume":"10","author":"Langmead","year":"2009","unstructured":"Langmead, B., Trapnell, C., Pop, M., & Salzberg, S. L. Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. 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