|ApexPrep DNA Plasmid Miniprep Kit
used for isolation of plasmid DNA
Sample solution is provided at 25 µL, 10mM.
For purification of up to 20 μg molecular biology grade plasmid DNA
- Ready-to-use plasmid DNA in minutes
- Reproducible yields of molecular biology grade plasmid DNA
- Single protocol for high- and low-copy vectors
The ApexPrep Spin Miniprep Kit is designed for isolation of up to 20 μg high-purity plasmid or cosmid DNA for use in routine molecular biology applications such as fluorescent and radioactive sequencing and cloning.
DNA Miniprep Protocol Using a Microcentrifuge
1. Inoculate 1-5 ml LB medium (selective antibiotic added) with a single colony, incubate with vigorous shaking at 37 °C overnight.
2. Harvest bacteria by centrifuging at > 6000 rpm for 5 min. Remove supernatant and keep pellet.
3. Resuspend bacterial pellet with 250 μl Buffer P1 and transfer to a fresh microcentrifuge tube.
4. Add 250 μl Buffer P2 and mix gently by inverting 5-6 times at room temperature.
5. Add 350 μl Buffer P3 and mix gently by inverting 5-6 times. Spin down at 12000 rpm for 10 min to remove bacterial/protein debris. Transfer supernatant to a spin column.
6. Centrifugation at 6000 rpm for 1 min. Discard flow-through.
7. (Optional), wash spin column with 0.5 ml Buffer PB; centrifuge at 6000 rpm for 1 min, and discard flow-through.
8. Wash spin column with 0.75 ml Buffer PE; centrifuge at 6000 rpm for 1 min, and discard flow-through.
9. Centrifuge at 12000 rpm for an additional 1 min to remove residual wash buffer.
10. To elute DNA, place the column in a clean 1.5 ml collection tube. Add 50 μl Buffer EB to the center of the membrane, let stand for 1 min, and centrifuge at 12000 rpm for 1 min to collect eluted DNA.