EndoFree Plasmid Mini Kit
2024-12-17Plasmid Mid Kit Liquid Form
2024-12-17Product Number: PLK0601
Shipping and Storage
- When using for the first time, add all RNase A carried by the reagent kit to Buffer YP1 (final concentration 100μg/ml) and store at 4℃. If RNase A is inactivated in Buffer YP1, there may be trace RNA residues in the extracted plasmid. Adding RNase A to Buffer YP1 is sufficient.
- SDS in Buffer YP2 may precipitate turbidity or precipitate when the ambient temperature is low. 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 reducing activity and facilitate transportation, Lyticase (2500U) is provided as a freeze-dried powder. After receiving it, it can be briefly centrifuged and dissolved in 0.25ml of sterilized water to prepare 10U/Ur. As repeated freeze-thaw cycles may reduce enzyme activity, it should be immediately packaged and stored according to the amount used each time, and stored at -20℃.
- 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 | PLK0601 50 Preps |
| RNase A (10mg/m1) | -20℃ | 150μl |
| Lyticase | -20℃ | 2500U |
| Buffer YP1 | 4℃ | 15 ml |
| Buffer YP2 | RT | 15 ml |
| Buffer YP3 | RT | 20 ml |
| Buffer PD | RT | 25 ml |
| Buffer WB | RT | 15 ml |
| Buffer EB | RT | 15 ml |
| Adsorption column AC | RT | 50 |
| Collection tube (2ml) | RT | 50 |
Description
This reagent kit uses an improved SDS alkaline lysis method to lyse cells and combines lysase specific digestion of yeast cell walls to isolate high-purity plasmid DNA from yeast culture medium within 1 hour. After yeast collection, wall breaking enzymes are added to remove the cell wall, followed by alkaline lysis of the cells. The silica matrix membrane in the centrifuge adsorption column selectively binds to plasmid DNA in the solution under high salt and low pH conditions. Impurities and other bacterial components are then removed through Buffer PD and Buffer WB. Finally, pure plasmid DNA is washed off the silica matrix membrane using low salt and high pH Buffer EB.
Features
- The silicon matrix membrane inside the centrifugal adsorption column is entirely made of specially designed adsorption membranes from world-renowned imported companies, with minimal differences in adsorption capacity between columns and good repeatability. Overcoming the drawback of unstable membrane quality in domestic reagent kits.
- Fast and convenient, without the need for toxic reagents such as phenol and chloroform, and without the need for ethanol precipitation. The obtained plasmids have high yield and good purity, and can be directly used for various molecular biology experiments such as enzyme digestion, transformation, PCR, in vitro transcription, sequencing, etc.
Application
Suitable for small-scale plasmid preparation in yeast
