Detection and enumeration of Lak megaphages in microbiome samples by endpoint and quantitative PCR
Summary: Lak megaphages are prevalent across diverse gut microbiomes and may potentially impact animal and human health through lysis of Prevotella. Given their large genome size (up to 660 kbp), Lak megaphages are difficult to culture, and their identification relies on molecular techniques. Here,...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721007358 |
| _version_ | 1830229882822459392 |
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| author | Marco A. Crisci Paula M. Corsini Nicola Bordin Lin-Xing Chen Jillain F. Banfield Joanne M. Santini |
| author_facet | Marco A. Crisci Paula M. Corsini Nicola Bordin Lin-Xing Chen Jillain F. Banfield Joanne M. Santini |
| author_sort | Marco A. Crisci |
| collection | DOAJ |
| description | Summary: Lak megaphages are prevalent across diverse gut microbiomes and may potentially impact animal and human health through lysis of Prevotella. Given their large genome size (up to 660 kbp), Lak megaphages are difficult to culture, and their identification relies on molecular techniques. Here, we present optimized protocols for identifying Lak phages in various microbiome samples, including procedures for DNA extraction, followed by detection and quantification of genes encoding Lak structural proteins using diagnostic endpoint and SYBR green-based quantitative PCR, respectively.For complete details on the use and execution of this protocol, please refer to Crisci et al., (2021). |
| first_indexed | 2024-12-18T10:42:42Z |
| format | Article |
| id | doaj.art-64411152fcab4303927f2b910283004e |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-18T10:42:42Z |
| publishDate | 2022-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-64411152fcab4303927f2b910283004e2022-12-21T21:10:36ZengElsevierSTAR Protocols2666-16672022-03-0131101029Detection and enumeration of Lak megaphages in microbiome samples by endpoint and quantitative PCRMarco A. Crisci0Paula M. Corsini1Nicola Bordin2Lin-Xing Chen3Jillain F. Banfield4Joanne M. Santini5Department of Structural and Molecular Biology, Division of Biosciences, University College London, London, UK; Corresponding authorQuadram Institute Bioscience, Norwich Research Park, Norwich, UKDepartment of Structural and Molecular Biology, Division of Biosciences, University College London, London, UKDepartment of Earth and Planetary Science, University of California Berkeley, Berkeley, CA, USA; Innovative Genomics Institute, University of California Berkeley, Berkeley, CA, USADepartment of Earth and Planetary Science, University of California Berkeley, Berkeley, CA, USA; Innovative Genomics Institute, University of California Berkeley, Berkeley, CA, USA; The University of Melbourne, Melbourne, VIC, Australia; Corresponding authorDepartment of Structural and Molecular Biology, Division of Biosciences, University College London, London, UK; Corresponding authorSummary: Lak megaphages are prevalent across diverse gut microbiomes and may potentially impact animal and human health through lysis of Prevotella. Given their large genome size (up to 660 kbp), Lak megaphages are difficult to culture, and their identification relies on molecular techniques. Here, we present optimized protocols for identifying Lak phages in various microbiome samples, including procedures for DNA extraction, followed by detection and quantification of genes encoding Lak structural proteins using diagnostic endpoint and SYBR green-based quantitative PCR, respectively.For complete details on the use and execution of this protocol, please refer to Crisci et al., (2021).http://www.sciencedirect.com/science/article/pii/S2666166721007358Health SciencesImmunologyMicrobiologyMolecular Biology |
| spellingShingle | Marco A. Crisci Paula M. Corsini Nicola Bordin Lin-Xing Chen Jillain F. Banfield Joanne M. Santini Detection and enumeration of Lak megaphages in microbiome samples by endpoint and quantitative PCR STAR Protocols Health Sciences Immunology Microbiology Molecular Biology |
| title | Detection and enumeration of Lak megaphages in microbiome samples by endpoint and quantitative PCR |
| title_full | Detection and enumeration of Lak megaphages in microbiome samples by endpoint and quantitative PCR |
| title_fullStr | Detection and enumeration of Lak megaphages in microbiome samples by endpoint and quantitative PCR |
| title_full_unstemmed | Detection and enumeration of Lak megaphages in microbiome samples by endpoint and quantitative PCR |
| title_short | Detection and enumeration of Lak megaphages in microbiome samples by endpoint and quantitative PCR |
| title_sort | detection and enumeration of lak megaphages in microbiome samples by endpoint and quantitative pcr |
| topic | Health Sciences Immunology Microbiology Molecular Biology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721007358 |
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