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.

已经开发了多种技术，包括 CRISPR-Cas9 基因组编辑，以生产转基因细胞系和动物模型。在许多情况下，基因组编辑技术的成功取决于引入的 DNA 的质量。然而，尚未探索小规模显微注射所需的高质量质粒的制备。在这里，我们比较了各种类型的质粒制备方法的显微注射能力，并开发了一种适用于秀丽隐杆线虫的高效且经济实惠的质粒微型制备方法显微注射。通过结合 Triton X-114 和基于柱的微型制备（因此，我们将其命名为 TXC）的优势，新的 TXC 方法经济实惠、高效，并且与基于显微注射效率的昂贵质粒中提方法相当。此外，TXC与一般分子生物学级反应兼容，能熟练处理不同类型的质粒。