A variety of techniques, including CRISPR-Cas9 genome editing, have been developed to produce genetically modified cell lines and animal models. In many cases, the success of the genome-editing techniques is dependent on the quality of the introduced DNA. However, the preparation of high-quality plasmids required for small-scale microinjection has not been explored. Here, we compared various types of plasmid preparation methods for their microinjection proficiency and developed an efficient and affordable plasmid mini preparation method suitable for Caenorhabditis elegans microinjection. By combining the advantages of Triton X-114 and column-based mini preparation (hence, we named it TXC), the new TXC method was affordable, efficient, and equivalent to expensive plasmid midiprep method based on microinjection efficiency. Besides, TXC was compatible with general molecular biology grade reactions and worked proficiently for different types of plasmids.
Keywords: CRISPR-Cas9; Genome editing; Microinjection; Plasmid miniprep.
© King Abdulaziz City for Science and Technology 2020.