Evaluation
Components of the kit were tested for isolation of plasmid DNA from bacterial cells according to the protocol
Stability
Cell pellets should yield plasmid DNA till the end of the expiration date
A260/A280
1.7 ~ 1.9
Description
The overall goal is to isolate extra-chromosomal, plasmid DNA from bacterial cells that contain the plasmid. The cells are first re-suspended in a suitable buffer (that minimizes nuclease activity) and ruptured to release the cellular contents. Subsequently, the plasmid DNA and RNA are separated from genomic DNA. Finally, by decreasing the “effective concentration” of water, the plasmid DNA is precipitated through sheer desolvation effects. If desired, the contaminating RNA can be removed by digestion with RNase A.
Includes
| Components | 15 Experiments | 25 Experiments |
|---|
| Bacterial Cell Pellet | 18 vials | 28 vials |
| Solution I | 3 ml | 5 ml |
| 5 N NaOH | 400 mcl | 750 mcl |
| 10% SDS | 1 ml | 3 ml |
| Sterile water | 10 ml | 30 ml |
| Solution III | 4 ml | 10 ml |
| DNA precipitation solution | 14 ml | 25 ml |
| Wash solution | 14 ml | 25 ml |
| TE buffer | 500 mcl | 750 mcl |
| Gel loading dye | 100 mcl | 200 mcl |
| Agarose | 1.5 gm | 2.5 gm |
| Ethidium bromide | 30 mcl | 75 mcl |
| 50X TAE buffer | 25 ml | 50 ml |
| Plasmid control | 60 mcl | 100 mcl |
