Endotoxin is a common contaminant in plasmid extraction. Because eukaryotic cells are very sensitive to endotoxin, the transfection efficiency will be greatly reduced if endotoxin is contained in the plasmid. This kit provides a simple, rapid and efficient method for the extraction of endotoxin-free plasmids. Based on conventional alkaline lysis method, the new and unique silicon membrane binds plasmid DNA efficiently and specifically. At the same time by using a special buffer system and endotoxin-removal filters, impurities such as endotoxin, genomic DNA, RNA, proteins are effectively removed.100-300 ml of bacterial culture can be processed each time, and up to 2 mg of transfection-grade plasmid DNA can be obtained. The entire extraction process takes only 50 minutes. The plasmids obtained from the kit have high purity and high yield and are particularly suitable for transfection experiments. The plasmids can also be used for DNA sequencing, PCR, in vitro transcription, restriction endonuclease digestion and other experiments.