Work

Simple and Fail-safe Method to Transform Miniprep Escherichia coli Strain K12 Plasmid DNA Into Viable Agrobacterium tumefaciens EHA105 Cells for Plant Genetic Transformation

Year: 2024

Type: article

Abstract: cells, making it a reproducible and accessible protocol for various genetic engineering applications. The transformation efficiency is consistently high, enhancing plasmid uptake while maintaining cell viability, requiring minimal specialized equipment and reagents. The proposed protocol offers significant advantages, including simplicity, reliability, and cost-effectiveness, positioning it as a valuable alternative to traditional techniques in the field of plant biotechnology. Key features • Uses liquid nitrogen as a proxy for freezing. • Plasmid DNA from competent bacterial cells is extracted using a user-friendly high-copy isolation kit. • A maximum of five consecutive days is sufficient to complete the procedures.

Source: BIO-PROTOCOL

DOI: 10.21769/bioprotoc.5174

Authors: Beenzu Siamalube, Emmanuel Ehinmitan, Maina Ngotho, Justus Onguso, Steven Runo

Institutions: Pan African University Institute for Basic Sciences, Technology and Innovation, Jomo Kenyatta University of Agriculture and Technology, Kenyatta University

Language: English

Topic: Plant tissue culture and regeneration

Subfield: Plant Science

Field: Agricultural and Biological Sciences

Domain: Life Sciences

SDG: Zero hunger

Open Access status: diamond

Funders: African Union, African Union Commission