Extraction of Geminivirus and Associated Satellite DNA from Leaf Samples.

Methods Mol Biol

Instituto de Hortofruticultura Subtropical y Mediterránea 'La Mayora' (IHSM-UMA-CSIC), Consejo Superior de Investigaciones Científicas, Algarrobo-Costa, Málaga, Spain.

Published: March 2025

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.

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http://dx.doi.org/10.1007/978-1-0716-4454-6_1DOI Listing

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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.

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