Method for recovery of intact DNA for community analysis of marine intertidal microbial biofilms
A protocol is described for rapid DNA isolation from marine biofilm microorganisms embedded in large amounts of exopolysaccharides. The method is a modification of the hot phenol protocol used for plants tissues, where nonexpensive and easily available enzymes were used. The method is based on the i...
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| Veröffentlicht in: | Molecular biotechnology 2005-05, Vol.30 (1), p.51-56 |
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| creator | Narváez-Zapata, José A Rodríguez-Avila, Norma Ortega-Morales, Benjamín O |
| description | A protocol is described for rapid DNA isolation from marine biofilm microorganisms embedded in large amounts of exopolysaccharides. The method is a modification of the hot phenol protocol used for plants tissues, where nonexpensive and easily available enzymes were used. The method is based on the incubation of biofilm biomass samples in an extraction buffer mixed with phenol preheated at 65 degrees C. The procedure can be completed in 2 h and up to 20 samples can be processed simultaneously with ease and DNA of excellent quality, as shown by successfully amplification of polymerase chain reaction (PCR) products. DNA was recovered from a range of intertidal marine biofilms with varying amounts of exopolysaccharides. |
| doi_str_mv | 10.1385/mb:30:1:051 |
| format | Article |
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The method is a modification of the hot phenol protocol used for plants tissues, where nonexpensive and easily available enzymes were used. The method is based on the incubation of biofilm biomass samples in an extraction buffer mixed with phenol preheated at 65 degrees C. The procedure can be completed in 2 h and up to 20 samples can be processed simultaneously with ease and DNA of excellent quality, as shown by successfully amplification of polymerase chain reaction (PCR) products. DNA was recovered from a range of intertidal marine biofilms with varying amounts of exopolysaccharides.</description><identifier>ISSN: 1073-6085</identifier><identifier>EISSN: 1073-6085</identifier><identifier>EISSN: 1559-0305</identifier><identifier>DOI: 10.1385/mb:30:1:051</identifier><identifier>PMID: 15805576</identifier><language>eng</language><publisher>United States: Springer Nature B.V</publisher><subject>Bacteria ; Bacteria - classification ; Bacteria - genetics ; Bacterial Typing Techniques - methods ; Biofilms ; Biofilms - classification ; Deoxyribonucleic acid ; DNA ; DNA, Bacterial - genetics ; DNA, Bacterial - isolation & purification ; Marine biology ; Microorganisms ; Oceans and Seas ; Phenols ; Polymerase Chain Reaction - methods ; Studies ; Water Microbiology</subject><ispartof>Molecular biotechnology, 2005-05, Vol.30 (1), p.51-56</ispartof><rights>Humana Press Inc. 2005</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c377t-7650ef71e128cce38c630b27df97c43578ac01f0240793d86c4e26c318be09d83</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,781,785,27928,27929</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15805576$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Narváez-Zapata, José A</creatorcontrib><creatorcontrib>Rodríguez-Avila, Norma</creatorcontrib><creatorcontrib>Ortega-Morales, Benjamín O</creatorcontrib><title>Method for recovery of intact DNA for community analysis of marine intertidal microbial biofilms</title><title>Molecular biotechnology</title><addtitle>Mol Biotechnol</addtitle><description>A protocol is described for rapid DNA isolation from marine biofilm microorganisms embedded in large amounts of exopolysaccharides. 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| ispartof | Molecular biotechnology, 2005-05, Vol.30 (1), p.51-56 |
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| subjects | Bacteria Bacteria - classification Bacteria - genetics Bacterial Typing Techniques - methods Biofilms Biofilms - classification Deoxyribonucleic acid DNA DNA, Bacterial - genetics DNA, Bacterial - isolation & purification Marine biology Microorganisms Oceans and Seas Phenols Polymerase Chain Reaction - methods Studies Water Microbiology |
| title | Method for recovery of intact DNA for community analysis of marine intertidal microbial biofilms |
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