Genenmed® Plasmid Kit provides an easy and rapid method for purification of the plasmid DNA from bacterial cells. The entire procedure takes less than 30min and multiple samples can be processed simultaneously. Genenmed® Plasmid Kit is optimized for purifying up to 30ug of the plasmid DNA. The purified plasmid DNA is ready for various further works, including cell transfection, sequencing, PCR, restriction digestion and electroporation without any additional manipulations.
Principle
Genenmed® Plasmid Kit utilizes a combination of the modified alkaline lysis method and the glass microfiber membrane, to which the plasmid DNA binds in the presence of chaotropic salts. The bound plasmid DNA is washed and eluted with ddH2O or TE buffer. This simple method eliminates the need for organic solvent extractions and alcohol precipitation.
Kit Contents
Keep Buffer PA at 4℃ and all other kit components at room temperature.
Before use, add 52ml absolute ethanol to each bottle of the Buffer PW.
Number of Preps
200
Spin columns
200
Buffer PA
55ml
Buffer PB
55ml
Buffer PC
75ml
Buffer PW (conc.)
13ml (each bottle)
Chemical Hazard
Buffer PC contains an irritant and can be harmful when contacted with skin, or when inhaled or swallowed. Care should be taken during handling. Always gloves and eye protector, and follow standard safety precautions.
Genenmed® Plasmid Kit Protocol
Perform centrifugation at full speed (>10,000Xg or 10,000~13,000rpm)
1. Pellet the 1~5ml saturated bacterial culture by centrifugation
2. Resuspend the bacterial pellet using 250ul Buffer PA
3. Transfer the suspension to a new 1.5ml tube
4. Add 250ul Buffer PB and mix completely by inverting the tube (do not vortex)
5. Incubate for 1~5min until the cell suspension becomes clear and viscous
6. Add 350ul Buffer PC and mix by inverting the tube 4~6 times (do not vortex)
7. Centrifuge for 10min
8. Transfer the supernatant to a spin column and centrifuge for 1min
9. Pour off the flow-through and re-insert the spin column to the collection tube
10. Apply 600ul Buffer PW and centrifuge for 1min
11. Pour off the flow-through and re-insert the spin column to the collection tube
12. Centrifuge for 3min to completely remove residual Buffer PW
13. Remove spin column from the collection tube and air-dry for 5min
14. Re-insert the spin column to a new 1.5ml tube with no cap
15. Add 50~100ul ddH2O or TE buffer (pH 8.0) and let stand for 1min
16. Centrifuge for 1min
17. Transfer the eluted plasmid DNA to a new 1.5ml tube
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