Agarose gel electrophoresis
Extracted Plasmid DNA is compared with Plasmid Control
A260/A280
1.6-1.9
Absence of nucleases
None detected
Stability
Stable for one year
Description
SpinCol Plasmid Miniprep Kit is designed for rapid and cost-effective small-scale preparation of high quality plasmid DNA from recombinant E.coli cultures. The kit utilizes an exclusive silica-based membrane technology in the form of a convenient spin column. The actual plasmid yield and optimal culture volume depend on the plasmid copy number and medium used for cultivation. Pelleted bacterial cells are resuspended and subjected to SDS/alkaline lysis to liberate the plasmid DNA. The resulting lysate is neutralized to create appropriate conditions for binding of plasmid DNA on the silica membrane in the spin column. Cell debris and SDS precipitate are pelleted by centrifugation, and the supernatant containing the plasmid DNA is loaded onto the spin column membrane. The adsorbed DNA is washed to remove contaminants, and is then eluted with a small volume of the Elution Buffer (10 mM Tris-HCl, pH 8.5).The RNA is removed when the cell pellets are resuspended in the resuspension buffer containing RNaseA.
Includes
| Components | 20 Experiments | 50 Experiments |
|---|
| Spin Column | 20 Nos | 50 Nos |
| Resuspension Solution | 6 ml | 14 ml |
| Lysis Solution | 6 ml | 14 ml |
| Neutralizing solution | 8 ml | 20 ml |
| Binding Buffer | 12 ml | 28 ml |
| Wash Buffer | 25 ml | 60 ml |
| Elution Buffer | 1.0 ml | 2.0 ml |
| Cell pellets (3ml) | 20 Nos | 50 Nos |
| Rnase A (10mg/ml) | 240 mcl | 600 mcl |
| Plasmid Control | 80 mcl | 150 mcl |
| 6x Gel loading dye | 60 mcl | 120 mcl |
| Ethidium Bromide | 50 mcl | 250 mcl |
| Agarose | 1.5 g | 3.0 g |
| 50X TAE Buffer | 25 ml | 50 ml |
