Vader-­Trap High-­Purity Plasmid DNA Purification Kit

Virongy’s Vader-Trap High-Purity MiniSpin Plasmid Purification Kit is built upon our proprietary technology. Our Vader-Trap plasmid purification technology significantly improves the traditional alkaline lysis procedure for plasmid purification. Virongy’s Vader-Trap technology uses innovative absorption particles and reagents to adsorb and remove contaminants and impurities, resulting in higher yield and greater purity than conventional miniprep spin kits. The purified plasmid DNA is suitable for many downstream applications, such as restriction digestion, transfection, cloning, and DNA sequencing, all with greater efficiency.

Highlights

Vader-Trap technology adsorbs and removes impurities, resulting in:

• Higher purity
• Higher yield
• Longer DNA sequencing read

Higher Yield

Higher plasmid DNA yields than those from other commercial MiniSpin kits (e.g., commercial Q kit).

Higher Purity

Higher plasmid DNA purity than from other commercial MiniSpin kits (e.g., commercial Q kit).

High-Quality DNA for Transfection

High quality plasmid DNA that can be directly used for transfection of cells.

Longer DNA Sequencing Reads

High-quality plasmid DNA that can be directly used for DNA sequencing.

Kit Components:

• Buffer VT
• Buffer A
• Buffer B
• Buffer C
• Buffer D
• Buffer E
• RNase A
• Spin Column Assembly & Collection Tubes

Protocol

Before You Start

• • Add RNase A to Buffer A. Store at 4^oC.
  • Add 10 ml of 100% ethanol to Buffer C.
  • Add 25 ml of 100% ethanol to Buffer D.

This protocol is designed for purification of up to 30 µg of plasmid DNA from 1.5 – 3 ml overnight cultures of E. coli.

• • 1. Take 1.5 ml of an overnight bacterial culture and place into a microcentrifuge tube that holds at least 1.8 ml.
    2. Resuspend particles in Buffer VT bottle by vigorously shaking. Take 150 μl of Buffer VT, add to the 1.5 ml bacterial culture tube, and mix them by inverting the tube 4-5 times.
    3. Centrifuge the bacterial culture/Buffer VT mixture at maximum speed (13,000-16,000 rpm) for 60 seconds. Carefully discard the supernatant without disturbing the pellet.
    4. Resuspend the pellet by adding 150 μl of Buffer A into the microcentrifuge tube and pipetting vigorously.
    5. Add 150 μl of Buffer B to the resuspended pellet mix, and lyse bacterial cells by inverting the tube 4-5 times.
    6. Neutralize the lysis reaction by adding 600 μl of Buffer C to the microcentrifuge tube and inverting the tube 4-5 times.
    7. Centrifuge at maximum speed for 10 minutes. A compact black pellet will form.
    8. Decant the supernatant, which contains the plasmid DNA, into a spin column assembly. Centrifuge for 30 seconds and discard flow-through. The plasmid DNA will bind to the column.
    9. Add 650 μl of Buffer D to the spin column to wash away contaminants. Centrifuge for 30 seconds at maximum speed. Discard the flow-through.
    10. Remove any residual buffer by centrifuging the spin column at maximum speed for 30 seconds. Discard the flow-through.
    11. Elute plasmid DNA by placing the spin column into a clean 1.5-ml collection tube (provided) and adding 50 μl of Buffer E. Incubate for 60 seconds at room temperature. Then centrifuge at maximum speed for 60 seconds.
    12. Store the eluted plasmid DNA at 4^o

References

Birnboim, H.C., & Doly, J. (1979). A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research, 7(6), 1513-1523.