Rapid isolation of fungal genomic DNA suitable for long distance PCR
A quick and reliable method for screening fungal transformants for specific genetic modifications is essential for many molecular applications. We have compared the applicability of a few rapid DNA extraction methods for Myrothecium and Aspergillus and tested the resulting DNA as to its suitability...
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| Veröffentlicht in: | Biotechnology letters 2007-12, Vol.29 (12), p.1845-1855 |
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| creator | De Maeseneire, S. L Van Bogaert, I. N Dauvrin, T Soetaert, W. K Vandamme, E. J |
| description | A quick and reliable method for screening fungal transformants for specific genetic modifications is essential for many molecular applications. We have compared the applicability of a few rapid DNA extraction methods for Myrothecium and Aspergillus and tested the resulting DNA as to its suitability for PCR. For Myrothecium gramineum, the highest DNA concentration was obtained with the procedure described by N. Vanittanakom et al. (J Clin Microbiol 2002, 40: 1739-1742). For A. nidulans, concentrations higher than 100 ng/μl were reached with the glass bead, the LiCl, the boiling, the liquid N₂ and the protoplast-based method. Samples of M. gramineum resulting from the boiling and the liquid N₂ procedure were suitable for the amplification of fragments up to 2.3 kb. The direct use of mycelium from M. gramineum in the PCR tube can be employed for the reproducible amplification of fragments up to 1 kb. Amplification of fragments up to 4.3 kb requires the use of the Elongase Mix on samples extracted with the liquid N₂ procedure. |
| doi_str_mv | 10.1007/s10529-007-9483-6 |
| format | Article |
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L ; Van Bogaert, I. N ; Dauvrin, T ; Soetaert, W. K ; Vandamme, E. J</creator><creatorcontrib>De Maeseneire, S. L ; Van Bogaert, I. N ; Dauvrin, T ; Soetaert, W. K ; Vandamme, E. J</creatorcontrib><description>A quick and reliable method for screening fungal transformants for specific genetic modifications is essential for many molecular applications. We have compared the applicability of a few rapid DNA extraction methods for Myrothecium and Aspergillus and tested the resulting DNA as to its suitability for PCR. For Myrothecium gramineum, the highest DNA concentration was obtained with the procedure described by N. Vanittanakom et al. (J Clin Microbiol 2002, 40: 1739-1742). For A. nidulans, concentrations higher than 100 ng/μl were reached with the glass bead, the LiCl, the boiling, the liquid N₂ and the protoplast-based method. Samples of M. gramineum resulting from the boiling and the liquid N₂ procedure were suitable for the amplification of fragments up to 2.3 kb. The direct use of mycelium from M. gramineum in the PCR tube can be employed for the reproducible amplification of fragments up to 1 kb. Amplification of fragments up to 4.3 kb requires the use of the Elongase Mix on samples extracted with the liquid N₂ procedure.</description><identifier>ISSN: 0141-5492</identifier><identifier>EISSN: 1573-6776</identifier><identifier>DOI: 10.1007/s10529-007-9483-6</identifier><identifier>PMID: 17680211</identifier><identifier>CODEN: BILED3</identifier><language>eng</language><publisher>Dordrecht: Dordrecht : Springer Netherlands</publisher><subject>Aspergillus nidulans - genetics ; Biological and medical sciences ; Biotechnology ; Deoxyribonucleic acid ; DNA ; DNA, Fungal - isolation & purification ; Electrophoresis, Agar Gel ; Fundamental and applied biological sciences. Psychology ; Genome, Fungal ; Genomic DNA isolation ; Long distance PCR ; Mycelium - genetics ; Myrothecium gramineum ; Polymerase Chain Reaction - methods</subject><ispartof>Biotechnology letters, 2007-12, Vol.29 (12), p.1845-1855</ispartof><rights>2008 INIST-CNRS</rights><rights>Springer Science+Business Media B.V. 2007</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c380t-3f716eac88a8d9fe290c58a4adb306b2eddb6ee8f4450cab0071a4b5428e96583</citedby><cites>FETCH-LOGICAL-c380t-3f716eac88a8d9fe290c58a4adb306b2eddb6ee8f4450cab0071a4b5428e96583</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19373664$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17680211$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>De Maeseneire, S. L</creatorcontrib><creatorcontrib>Van Bogaert, I. N</creatorcontrib><creatorcontrib>Dauvrin, T</creatorcontrib><creatorcontrib>Soetaert, W. K</creatorcontrib><creatorcontrib>Vandamme, E. J</creatorcontrib><title>Rapid isolation of fungal genomic DNA suitable for long distance PCR</title><title>Biotechnology letters</title><addtitle>Biotechnol Lett</addtitle><description>A quick and reliable method for screening fungal transformants for specific genetic modifications is essential for many molecular applications. We have compared the applicability of a few rapid DNA extraction methods for Myrothecium and Aspergillus and tested the resulting DNA as to its suitability for PCR. For Myrothecium gramineum, the highest DNA concentration was obtained with the procedure described by N. Vanittanakom et al. (J Clin Microbiol 2002, 40: 1739-1742). For A. nidulans, concentrations higher than 100 ng/μl were reached with the glass bead, the LiCl, the boiling, the liquid N₂ and the protoplast-based method. Samples of M. gramineum resulting from the boiling and the liquid N₂ procedure were suitable for the amplification of fragments up to 2.3 kb. The direct use of mycelium from M. gramineum in the PCR tube can be employed for the reproducible amplification of fragments up to 1 kb. Amplification of fragments up to 4.3 kb requires the use of the Elongase Mix on samples extracted with the liquid N₂ procedure.</description><subject>Aspergillus nidulans - genetics</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA, Fungal - isolation & purification</subject><subject>Electrophoresis, Agar Gel</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genome, Fungal</subject><subject>Genomic DNA isolation</subject><subject>Long distance PCR</subject><subject>Mycelium - genetics</subject><subject>Myrothecium gramineum</subject><subject>Polymerase Chain Reaction - methods</subject><issn>0141-5492</issn><issn>1573-6776</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNpd0MuKFDEUBuAgitOOPoAbDQO6Kz25p5ZDjzcYVEZnHU6lkiZDdaVNqha-vWm6YcBN8i--c0h-Ql4z-MAAzMfKQPG-a7HrpRWdfkI2TJkWjNFPyQaYZJ2SPb8gL2p9AIDegHlOLpjRFjhjG3Jzh4c00lTzhEvKM82RxnXe4UR3Yc775OnN92ta17TgMAUac6FTnnd0THXB2Qf6c3v3kjyLONXw6nxfkvvPn35vv3a3P758217fdl5YWDoRDdMBvbVoxz4G3oNXFiWOgwA98DCOgw7BRikVeBzavxjKQUluQ6-VFZfk_WnvoeQ_a6iL26fqwzThHPJanbZSMS6O8Oo_-JDXMre3OSOk1ZIp1hA7IV9yrSVEdyhpj-WvY-CO_bpTv-4Yj_063WbenBevwz6MjxPnQht4dwZYPU6xtI5SfXS9MEJr2dzbk4uYHe5KM_e_ODABYDlvp_gHHV-KIg</recordid><startdate>20071201</startdate><enddate>20071201</enddate><creator>De Maeseneire, S. 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Psychology</topic><topic>Genome, Fungal</topic><topic>Genomic DNA isolation</topic><topic>Long distance PCR</topic><topic>Mycelium - genetics</topic><topic>Myrothecium gramineum</topic><topic>Polymerase Chain Reaction - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>De Maeseneire, S. L</creatorcontrib><creatorcontrib>Van Bogaert, I. N</creatorcontrib><creatorcontrib>Dauvrin, T</creatorcontrib><creatorcontrib>Soetaert, W. K</creatorcontrib><creatorcontrib>Vandamme, E. 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L</au><au>Van Bogaert, I. N</au><au>Dauvrin, T</au><au>Soetaert, W. K</au><au>Vandamme, E. J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Rapid isolation of fungal genomic DNA suitable for long distance PCR</atitle><jtitle>Biotechnology letters</jtitle><addtitle>Biotechnol Lett</addtitle><date>2007-12-01</date><risdate>2007</risdate><volume>29</volume><issue>12</issue><spage>1845</spage><epage>1855</epage><pages>1845-1855</pages><issn>0141-5492</issn><eissn>1573-6776</eissn><coden>BILED3</coden><abstract>A quick and reliable method for screening fungal transformants for specific genetic modifications is essential for many molecular applications. We have compared the applicability of a few rapid DNA extraction methods for Myrothecium and Aspergillus and tested the resulting DNA as to its suitability for PCR. For Myrothecium gramineum, the highest DNA concentration was obtained with the procedure described by N. Vanittanakom et al. (J Clin Microbiol 2002, 40: 1739-1742). For A. nidulans, concentrations higher than 100 ng/μl were reached with the glass bead, the LiCl, the boiling, the liquid N₂ and the protoplast-based method. Samples of M. gramineum resulting from the boiling and the liquid N₂ procedure were suitable for the amplification of fragments up to 2.3 kb. The direct use of mycelium from M. gramineum in the PCR tube can be employed for the reproducible amplification of fragments up to 1 kb. Amplification of fragments up to 4.3 kb requires the use of the Elongase Mix on samples extracted with the liquid N₂ procedure.</abstract><cop>Dordrecht</cop><pub>Dordrecht : Springer Netherlands</pub><pmid>17680211</pmid><doi>10.1007/s10529-007-9483-6</doi><tpages>11</tpages></addata></record> |
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| ispartof | Biotechnology letters, 2007-12, Vol.29 (12), p.1845-1855 |
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| subjects | Aspergillus nidulans - genetics Biological and medical sciences Biotechnology Deoxyribonucleic acid DNA DNA, Fungal - isolation & purification Electrophoresis, Agar Gel Fundamental and applied biological sciences. Psychology Genome, Fungal Genomic DNA isolation Long distance PCR Mycelium - genetics Myrothecium gramineum Polymerase Chain Reaction - methods |
| title | Rapid isolation of fungal genomic DNA suitable for long distance PCR |
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