Abstract
Objective: To explore a simple, improved and inexpensive method for isolation of plasmid DNA and its value in molecular biological laboratory research. Methods: Plasmid DNA was isolated using general alkaline lysis method and improved method, respectively. The isolated plasmid DNA was quantified by spectrophotometric measurement, and evaluated by the electrophoresis of plasmid DNA in an agarose gel, the restriction enzyme digestion, and the transfection into eukaryotic cells. Results: The quantity of plasmid DNA isolated by the improved method was 3.2 μg per milliliter culture of transformed bacteria, and the ratio of A260/280 was 1.91. The plasmid DNA could be fully digested by restriction enzyme. The efficiency of the plasmid DNA transfection into eukaryotic cells by Lipofectamine2000 was about 20%. Conclusion: The improved method is simple, practical and economical. Plasmid DNA isolated by this method has been shown to be suitable for restriction enzyme digestion and cell transfection.
| Original language | English |
|---|---|
| Pages (from-to) | 106-108 |
| Number of pages | 3 |
| Journal | Journal of Xi'an Jiaotong University (Medical Sciences) |
| Volume | 28 |
| Issue number | 1 |
| State | Published - Feb 2007 |
Keywords
- Isolation
- Plasmid DNA
- Restriction enzyme digestion
- Transfection