Yeast Plasmid Mini Kit
2024-12-17EndoFree Plasmid Mid Scale Mini Kit
2024-12-17Product Number: PLK0801
Shipping and Storage
- When using for the first time, add all RNase A carried by the reagent kit to Buffer P1 (concentration 100μg/ml) and store at 4℃. If RNase A is inactivated in Buffer P1, the extracted plasmid may contain trace amounts of RNA residue. Adding RNase A to Buffer P1 is sufficient.
- Buffer ER can be stored at 4℃ for one month. If it needs to be stored for a long time, it is recommended to store it at -20℃! When the ambient temperature is low, SDS in Buffer P2 may precipitate and become turbid or precipitate. It can be heated in a 37℃ water bath for a few minutes to restore clarity. Do not shake violently to avoid excessive foam formation.
- To avoid volatilization, oxidation, and pH changes caused by prolonged exposure of reagents to the air, each solution should be covered tightly in a timely manner after use.
Components
| Component | Storage | PLK0801 20 Preps | PLK0802 40 Preps |
| RNase A(10mg/ml) | -20℃ | 0.75 ml | 1.3 ml |
| Buffer P1 | 4℃ | 65 ml | 130 ml |
| Buffer P2 | RT | 50 ml | 100 ml |
| Buffer P3 | RT | 50 ml | 110 ml |
| Buffer IRA | RT | 1.5 ml | 3 ml |
| Buffer IRB | RT | 15 ml | 30 ml |
| Buffer ER | -20℃ | 5 ml | 10 ml |
Description
This reagent kit extracts plasmid DNA from cultured bacteria using alkaline lysis method, using a unique solution formula and Buffer ER. It only requires a few simple centrifugations to remove impurities such as proteins, polysaccharides, endotoxins, RNA, etc., to obtain high-quality plasmid DNA. The OD260/280 of purified DNA is usually around 1.8, and the resulting plasmid can be directly applied in tasks that require high DNA purity, such as cell transfection and even animal in vivo experiments. The purification process in the later stage is operated in a 1.5ml small centrifuge tube, which is simple, does not require special equipment, does not require column passing, and does not require phenol chloroform extraction; Plasmids released by bacterial lysis can be completely recovered without worrying about the loss of plasmid DNA. This method extracts and purifies plasmid DNA with minimal damage to plasmids. Even large plasmids or ultra large BAC/PAC plasmids with a size of 10kb or even 100kb can be effectively purified as long as they can be extracted by alkaline lysis. You can choose to dissolve plasmids in any small volume, with a concentration of up to 5μg/μl. The super helix ratio can reach up to 95%, with no endotoxins and good transfection effect.
Features
- No toxic reagents such as phenol and chloroform are needed, and no ethanol precipitation is required. Fast and convenient extraction of 150μg-600μg pure high copy plasmid DNA from 50-70ml of Escherichia coli LB (Luria Bertani) culture medium, with an extraction rate of 80-90%.
- The obtained plasmids have high yield, concentration, super helix ratio, and purity, and can be directly used for various molecular biology experiments such as enzyme digestion, transformation, PCR, in vitro transcription, sequencing, etc.
- The endotoxin content is extremely low (<0.1EU/μg DNA) and can be directly applied to cell transfection.
Application
Suitable for medium to high purity or transfection level plasmid preparation and BAC/PAC large-scale plasmid preparation
