Plasmid DNA Miniprep Kit
No:DMDPN100 & 300
For research use only
Sample: up to 4 ml bacterial cells
Yield: up to 30μg of plasmid
Introduction
The BioDiamond Plasmid DNA Miniprep Kit provides a fast, simple, and cost-effective spin-column method for isolation of plasmid DNA from cultured bacterial cells. The BioDiamond Plasmid DNA Miniprep Kit is based on alkaline lysis of bacterial cells followed by binding of DNA onto the glass fiber matrix of the spin column in the presence of a high amount of salt. Phenol extraction and ethanol precipitation are not required, and high-quality plasmid DNA is eluted with a small volume of Tris buffer (included in each kit) or water (pH is between 7.0 and 8.5). Plasmid DNA purified with the BioDiamond Plasmid DNA Miniprep Kit is suitable for a variety of routine applications including restriction enzyme digestion, sequencing, library screening, in vitro translation, transfection of robust cells, ligation and transformation. The entire procedure can be completed within 20 minutes.
Kit Contents
No:DMDPN100 & 300
For research use only
Sample: up to 4 ml bacterial cells
Yield: up to 30μg of plasmid
Introduction
The BioDiamond Plasmid DNA Miniprep Kit provides a fast, simple, and cost-effective spin-column method for isolation of plasmid DNA from cultured bacterial cells. The BioDiamond Plasmid DNA Miniprep Kit is based on alkaline lysis of bacterial cells followed by binding of DNA onto the glass fiber matrix of the spin column in the presence of a high amount of salt. Phenol extraction and ethanol precipitation are not required, and high-quality plasmid DNA is eluted with a small volume of Tris buffer (included in each kit) or water (pH is between 7.0 and 8.5). Plasmid DNA purified with the BioDiamond Plasmid DNA Miniprep Kit is suitable for a variety of routine applications including restriction enzyme digestion, sequencing, library screening, in vitro translation, transfection of robust cells, ligation and transformation. The entire procedure can be completed within 20 minutes.
Kit Contents
NOTE
★ Add the provided RNase A solution to Buffer S1, mix, and store at 2–8°C.
★ Add ethanol (96–100%) to Buffer W2, shaking before use (see bottle label for volume).
★ Check Buffers before use for salt precipitation. Redissolve any precipitate by warming to 37°C.
★ Buffers S2, S3, and W1 contain irritants. Wear gloves when handling these buffers.
Quality Control
In accordance with FairBiotech’s ISO-certified Total Quality Management System, the quality of the BioDiamond
Plasmid DNA Miniprep Kit is tested on a lot-to-lot basis to ensure consistent product quality.
Additional requirements
*Ethanol (96~100%)
*1.5 ml microcentrifuge tubes
Protocol
Step 1 Bacterial Cells Harvesting
- Transfer 1.5 ml bacterial culture to a microcentrifuge tube
- Centrifuge at 14,000 x g for 1 minute and discard the supernatant.
- Resuspend pelleted bacterial cells in 200 μl Buffer S1 (RNase A added)
- Add 200 μl Buffer S2 and mix thoroughly by inverting the tube 10 times (Do not vortex) and then stand at room temperature for 2 minutes or until the lysate is homologous.
- Add 300 μl Buffer S3 and mix immediately and thoroughly by inverting the tube 10 times (Do not vortex). Centrifuge at 14,000 x g for 3 minutes.
- Place a SP Column in a Collection Tube. Apply the supernatant (from step 4) to the SP column by decanting or pipetting.
- Centrifuge at 14,000 x g for 30 seconds. Discard the flow-through and place the SP column back into the same Collection Tube.
- Add 400 µl of Buffer W1 into the SP Column. Centrifuge at 14,000 x g for 30 seconds. Discard the flow-through and place the SP column back into the same Collection tube.
- Add 600 µl of Buffer W2 (Ethanol added) into the SP Column. Centrifuge at 14,000 x g for 30 seconds. Discard the flow-through and place the SP column back into the same Collection tube.
- Centrifuge at 14,000 x g again for 2 minutes to remove residual Buffer W2.
- To elute DNA, place the SP column in a clean 1.5 ml microcentrifuge tube.
- Add 50-200 μl Buffer EL or water (pH is between 7.0 and 8.5) to the center of each SP column, let stand for 2 minutes, and centrifuge at 14,000 x g for 2 minutes.
Troubleshooting
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