Extraction of Geminivirus and Associated Satellite DNA from Leaf Samples

The isolation of DNA from geminiviruses and their associated DNA satellites in plant leaf samples is a key step, as high-quality DNA preparations are essential for studies related to the characterization of geminivirus genomes and their interactions with host plants. The protocol outlined in this ch...

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Detalhes bibliográficos
Autores: Fiallo-Olivé, Elvira, Mohammed, Heyam S, Zerbini, Francisco Murilo, Navas-Castillo, Jesús
Formato: otro
Estado:Versión aceptada para publicación
Fecha de publicación:2025
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/385403
Acesso em linha:http://hdl.handle.net/10261/385403
https://api.elsevier.com/content/abstract/scopus_id/105000082841
Access Level:acceso abierto
Palavra-chave:CTAB protocol
DNA satellites
Geminiviruses
Nucleic acid extraction
Descrição
Resumo:The isolation of DNA from geminiviruses and their associated DNA satellites in plant leaf samples is a key step, as high-quality DNA preparations are essential for studies related to the characterization of geminivirus genomes and their interactions with host plants. The protocol outlined in this chapter describes a methodology based on the use of cetyltrimethylammonium bromide (CTAB) that facilitates the efficient extraction of total nucleic acids from plant tissues. The DNA obtained through this protocol finds direct application in various molecular techniques such as polymerase chain reaction, Southern blot, dot blot, and rolling-circle amplification. Additionally, the obtained DNA remains stable for years when stored at -20 °C, even after periods at room temperature. In essence, this extraction protocol is simple and useful for both experienced researchers and those introducing themselves to the study of geminiviruses and DNA satellites.