Efficient protocols for the extraction of microbial DNA from the rhizosphere of hydrophilic forests in Chile

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dc.contributor.authorCaceres, Pablo
dc.contributor.authorCordero, Cecilia
dc.contributor.authorGonzalez, Gloria
dc.contributor.authorQuiroz, Karla
dc.contributor.authorBobadilla, Juan C.
dc.contributor.authorBravo, Carmen
dc.contributor.authorCaligari, Peter D. S.
dc.contributor.authorCarrasco, Basilio
dc.contributor.authorGarcia Gonzales, Rolando
dc.date.accessioned2024-01-10T12:38:38Z
dc.date.available2024-01-10T12:38:38Z
dc.date.issued2012
dc.description.abstractP. Caceres, C. Cordero, G. Gonzalez, K. Quiroz, J.C. Bobadilla, C. Bravo, P.D.S. Caligari, B. Carrasco, and R. Garcia-Gonzales. 2012. Efficient protocols for the extraction of microbial DNA from the rhizosphere of hydrophilic forests in Chile. Cien. Inv. Agr. 38(3): 585-592. A lysis buffer-based protocol (Protocol BA), a modified lysis buffer-based protocol (Protocol BA Mod) and a commercial extraction kit (Protocol PS Kit) (Power Soil, Mo bio Laboratories, CA USA) were each evaluated for their ability to produce high-quality DNA with yields sufficient to allow its use in biodiversity studies. Similarly, the effect of liquid nitrogen on the process of cell disruption in all of the protocols that were studied. DNA yields ranged from 12.4 ng g(-1) of processed soil to 9620 ng g(-1) using the modified lysis buffer and commercial extraction kit, respectively. The quality of the DNA was determined by the ability of the DNA to produce efficient and reproducible polymerase chain reaction (PCR) products, using primers for universal 16S and 18S ribosomal RNA regions from bacteria and fungi, respectively. High-quality DNA was obtained to run PCRs in all protocols, but the efficiency of the method depended on the dilution of the DNA prior to performing the PCR. The three extraction methods generated PCR products with 90% efficiency. The DNA produced with the commercial kit was able to produce the highest PCR efficiency (95%) when the 10(-1) dilution was used. The method based on the use of lysis buffer produced the highest efficiency (90%) using a 10(-2) dilution. Meanwhile, the modified lysis buffer-based protocol generated the highest efficiency of PCR products using the 10(-3) dilution factor with 95% of efficiency. For the first time, reliable and efficient DNA isolation from the rhizosphere of hydrolic forest is documented, enabling a wide range of applications for this technique.
dc.fechaingreso.objetodigital2024-05-30
dc.format.extent8 páginas
dc.fuente.origenWOS
dc.identifier.doi10.4067/S0718-16202012000300018
dc.identifier.issn0304-5609
dc.identifier.urihttps://doi.org/10.4067/S0718-16202012000300018
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/77077
dc.identifier.wosidWOS:000313400900018
dc.information.autorucAgronomía e Ing. Forestal;Carrasco B;S/I;174497
dc.issue.numero3
dc.language.isoen
dc.nota.accesoContenido completo
dc.pagina.final592
dc.pagina.inicio585
dc.publisherPONTIFICIA UNIV CATOLICA CHILE, FAC AGRONOMIA INGENIERIA FORESTAL
dc.revistaCIENCIA E INVESTIGACION AGRARIA
dc.rightsacceso abierto
dc.subjecthydrophilic forests
dc.subject16S ribosomal RNA
dc.subjectPCR
dc.subjectrhizosphere
dc.subjectsoil DNA extraction
dc.subjectPURIFICATION METHODS
dc.subjectSOIL
dc.subjectRECOVERY
dc.subjectTRUFFLE
dc.subjectRDNA
dc.subject.ods15 Life on Land
dc.subject.odspa15 Vida de ecosistemas terrestres
dc.titleEfficient protocols for the extraction of microbial DNA from the rhizosphere of hydrophilic forests in Chile
dc.typeartículo
dc.volumen39
sipa.codpersvinculados174497
sipa.indexWOS
sipa.indexScopus
sipa.trazabilidadCarga SIPA;09-01-2024
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