Yield: up to 30 ug of plasmid/cosmid DNA Format: spin column Operation time: 20 min Elution volume: 50-100 ul
High-Speed Plasmid Mini Kit
Sample: 1-4 ml of bacterial culture
Yield: up to 30 mg of plasmid/cosmid DNA
Format: spin column
Operation time: 20 min
Elution volume: 50-100 ml
Cat. No. S00101
Store at room temperature (15 - 25°C )
For research used only
IntroductionHigh-Speed Plasmid Mini Kit is designed for rapid isolation of plasmid or cosmid DNA from 1-4 ml of bacterial cultures. In the process, the modified alkaline lysis method (1) and RNase treatment are used to get cleared cell lysate with minimal genomic DNA and RNA contaminants. In the presence of a chaotropic salt, the plasmid DNA in the lysate binds to glass fiber matrix in the spin column (2). The contaminants are washed with an ethanol-contained wash buffer and finally, the purified plasmid DNA is eluted by low salt elution buffer or water. The protocol does not require DNA phenol extraction and alcohol precipitation. Typical yields are 10-20 mg for high-copy number plasmid or 0.5-5 mg for low-copy number plasmid. The entire procedure can be completed in 20 minutes and the purified plasmid DNA is ready for restriction digestion, ligation, PCR, and sequencing reaction.
The quality of High-Speed Plasmid Mini Kit is tested on a lot-to-lot basis. The Kits are tested by isolation of plasmid DNA from 4 ml culture of E. coli DH5α that contains the plasmid pBluescript (A600 > 2 units/ ml). More than 20 mg of plasmid DNA could be quantified with spectrophotometer. 1 mg of the purified plasmid is used on restriction enzyme digestion with Eco RI and digested DNA is checked by agarose gel analysis.
Kit ContentsName | S00101 | S00101-S |
PD1 Buffer* | 25 ml | 1 ml |
PD2 Buffer** | 25 ml | 1 ml |
PD3 Buffer | 45 ml | 1.5 ml |
W 1 Buffer | 45 ml | 2 ml |
Wash Buffer (concentrated)*** | 25 ml | 1 ml |
Elution Buffer (10mM Tris-HCl, pH 8.5 at 25°C) | 6 ml | 1 ml |
RNase A (50 mg/ ml) | 50 ml | - |
PD column | 100 pcs | 4 pcs |
2 ml Collection tube | 100 pcs | 4 pcs |
* Add provided RNase A to PD1 Buffer and store at 4°C.
** If precipitates have formed in PD2 Buffer, warm the buffer in a 37°C waterbath to dissolve precipitates.
*** Add 100 ml/ 4 ml ethanol (96-100%) to Wash Buffer prior to initial use.
The component contains irritant agent. During operation, always wear a lab coat, disposable gloves, and protective goggles.
References(1) Birnboim, H. C., and Doly, J. (1979) Nucleic Acids Res. 7, 1513.
(2) Vogelstein, B., and Gillespie, D. (1979) Proc. Natl. Acad. Sci. USA 76, 615.
High-Speed Plasmid Mini Kit
