Kim Hyun-Min, Tian Sisi, Wang Shouyue
School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, 300072 China.
3 Biotech. 2020 Aug;10(8):350. doi: 10.1007/s13205-020-02346-7. Epub 2020 Jul 22.
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 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.
已经开发出多种技术，包括CRISPR-Cas9基因组编辑技术，以产生基因编辑细胞系和动物模型。在许多情况下，基因组编辑技术的成功取决于导入DNA的质量。然而，尚未探索用于小规模显微注射所需的高质量质粒的制备方法。在这里，我们比较了各种类型的质粒制备方法在显微注射方面的熟练度，并开发了一种高效且经济实惠的适合显微注射的质粒小量制备方法。通过结合Triton X-114和基于柱的小量制备方法的优点（因此，我们将其命名为TXC），新的TXC方法经济实惠、效率高，并且基于显微注射效率与昂贵的质粒中量制备方法相当。此外，TXC与一般分子生物学级反应兼容，并且对不同类型的质粒都能有效发挥作用。
