Protocol for ex vivo competition and sequencing of mycobacterium isolated from infected guinea pigs
Summary: We describe steps for gDNA isolation from mycobacterium strains isolated from guinea pig lungs. We detail steps for infection of guinea pigs with Mycobacterium tuberculosis, followed by in vitro growth, gDNA isolation, and whole genome sequencing. We also describe an ex vivo competition exp...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722006840 |
| _version_ | 1828265394070093824 |
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| author | Saba Naz Shruti Dabral Dhiraj Kumar Vinay Kumar Nandicoori |
| author_facet | Saba Naz Shruti Dabral Dhiraj Kumar Vinay Kumar Nandicoori |
| author_sort | Saba Naz |
| collection | DOAJ |
| description | Summary: We describe steps for gDNA isolation from mycobacterium strains isolated from guinea pig lungs. We detail steps for infection of guinea pigs with Mycobacterium tuberculosis, followed by in vitro growth, gDNA isolation, and whole genome sequencing. We also describe an ex vivo competition experiment to determine the selective advantage of one strain over another. We include details for WGS and mutation spectrum analysis. The protocol can be used to identify mutations that arise in other pathogenic bacteria.For complete details on the use and execution of this protocol, please refer to Naz et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-13T04:38:41Z |
| format | Article |
| id | doaj.art-3d620ffe0dce46bb9a14bbf94e04785b |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-13T04:38:41Z |
| publishDate | 2022-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-3d620ffe0dce46bb9a14bbf94e04785b2022-12-22T03:02:05ZengElsevierSTAR Protocols2666-16672022-12-0134101804Protocol for ex vivo competition and sequencing of mycobacterium isolated from infected guinea pigsSaba Naz0Shruti Dabral1Dhiraj Kumar2Vinay Kumar Nandicoori3Mycobacterial Signalling Lab, CSIR-Centre for Cellular and Molecular Biology, Habsiguda, Telangana 500007, India; Signal Transduction Lab-I, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, Delhi 110067, India; Corresponding authorCellular Immunology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, Delhi 110067, IndiaCellular Immunology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, Delhi 110067, IndiaMycobacterial Signalling Lab, CSIR-Centre for Cellular and Molecular Biology, Habsiguda, Telangana 500007, India; Signal Transduction Lab-I, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, Delhi 110067, India; Corresponding authorSummary: We describe steps for gDNA isolation from mycobacterium strains isolated from guinea pig lungs. We detail steps for infection of guinea pigs with Mycobacterium tuberculosis, followed by in vitro growth, gDNA isolation, and whole genome sequencing. We also describe an ex vivo competition experiment to determine the selective advantage of one strain over another. We include details for WGS and mutation spectrum analysis. The protocol can be used to identify mutations that arise in other pathogenic bacteria.For complete details on the use and execution of this protocol, please refer to Naz et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166722006840GenomicsSequencingImmunologyMicrobiologyModel organisms |
| spellingShingle | Saba Naz Shruti Dabral Dhiraj Kumar Vinay Kumar Nandicoori Protocol for ex vivo competition and sequencing of mycobacterium isolated from infected guinea pigs STAR Protocols Genomics Sequencing Immunology Microbiology Model organisms |
| title | Protocol for ex vivo competition and sequencing of mycobacterium isolated from infected guinea pigs |
| title_full | Protocol for ex vivo competition and sequencing of mycobacterium isolated from infected guinea pigs |
| title_fullStr | Protocol for ex vivo competition and sequencing of mycobacterium isolated from infected guinea pigs |
| title_full_unstemmed | Protocol for ex vivo competition and sequencing of mycobacterium isolated from infected guinea pigs |
| title_short | Protocol for ex vivo competition and sequencing of mycobacterium isolated from infected guinea pigs |
| title_sort | protocol for ex vivo competition and sequencing of mycobacterium isolated from infected guinea pigs |
| topic | Genomics Sequencing Immunology Microbiology Model organisms |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722006840 |
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