{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T13:38:54Z","timestamp":1740145134055,"version":"3.37.3"},"reference-count":4,"publisher":"Cold Spring Harbor Laboratory","issue":"11","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cold Spring Harb Protoc"],"published-print":{"date-parts":[[2023,11]]},"abstract":"<jats:p>In this protocol, we describe the isolation of genomic DNA (gDNA) from<jats:italic>Staphylococcus aureus<\/jats:italic>strains using a chloroform extraction and ethanol precipitation method. This gDNA-isolation method is well-suited for downstream whole-genome sequencing applications when working with<jats:italic>S. aureus<\/jats:italic>strains that contain plasmids, as only a small amount of plasmid DNA is isolated along with the gDNA. Similar to other gDNA isolation methods for Gram-positive bacteria, the first step in the procedure is a mechanical lysis (e.g., using a bead beating grinder) or an enzymatic lysis step. In this protocol, the peptidoglycan layer of<jats:italic>S. aureus<\/jats:italic>is digested with an enzyme called lysostaphin. This enzyme cleaves pentaglycine cross-bridges within the peptidoglycan of<jats:italic>S. aureus.<\/jats:italic>After this lysis step, gDNA can be purified using similar procedures as those used for Gram-negative bacteria. We include additional cleanup and quantification procedures in the final steps of this protocol, in case the aim is to use the gDNA for genome-sequencing projects. By modifying the bacterial lysis step, the procedure can be easily adapted to isolate gDNA from other bacteria.<\/jats:p>","DOI":"10.1101\/pdb.prot107899","type":"journal-article","created":{"date-parts":[[2023,11,1]],"date-time":"2023-11-01T16:14:13Z","timestamp":1698855253000},"page":"pdb.prot107899","source":"Crossref","is-referenced-by-count":6,"title":["Preparation of<i>Staphylococcus aureus<\/i>Genomic DNA Using a Chloroform Extraction and Ethanol Precipitation Method, Followed by Additional Cleanup and Quantification Steps"],"prefix":"10.1101","volume":"2023","author":[{"given":"Merve S.","family":"Zeden","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6235-8687","authenticated-orcid":false,"given":"Angelika","family":"Gr\u00fcndling","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"246","published-online":{"date-parts":[[2023,4,28]]},"reference":[{"key":"2023110108200716000_2023.11.pdb.prot107899.1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(61)80047-8"},{"key":"2023110108200716000_2023.11.pdb.prot107899.2","doi-asserted-by":"publisher","DOI":"10.3791\/52134"},{"key":"2023110108200716000_2023.11.pdb.prot107899.3","doi-asserted-by":"publisher","DOI":"10.1101\/pdb.prot107897"},{"key":"2023110108200716000_2023.11.pdb.prot107899.4","doi-asserted-by":"publisher","DOI":"10.1101\/pdb.prot107900"}],"container-title":["Cold Spring Harbor Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/pdb.prot107899","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,1]],"date-time":"2023-11-01T16:14:27Z","timestamp":1698855267000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.cshprotocols.org\/lookup\/doi\/10.1101\/pdb.prot107899"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,4,28]]},"references-count":4,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2023,11,1]]},"published-print":{"date-parts":[[2023,11]]}},"alternative-id":["10.1101\/pdb.prot107899"],"URL":"https:\/\/doi.org\/10.1101\/pdb.prot107899","relation":{},"ISSN":["1940-3402","1559-6095"],"issn-type":[{"type":"print","value":"1940-3402"},{"type":"electronic","value":"1559-6095"}],"subject":[],"published":{"date-parts":[[2023,4,28]]}}}