AquaPlasmidTM is an aqueous solution-based reagent for purification of plasmid DNA. It extracts plasmid DNA without using alkaline lysis and without denaturing the plasmid DNA. It can yield an unmatched 10-15 ug of plasmid DNA from one milliliter of overnight bacterial culture. The extraction protocol consists of three simple steps: cell lysis, debris removal and DNA precipitation. However, if you like column-based purification, you may double its DNA yield by replacing the alkaline lysis solution in your kit with AquaPlasmid solution for cell lysis.
AquaPlasmid extracts plasmid DNA without using alkaline lysis and without denaturing the DNA.
The AquaPlasmid protocol consists of three simple steps: cell lysis, debris removal, and DNA precipitation.
AquaPlasmid is scalable. A single AquaPlasmid kit can be used for mini, midi, and maxi preps.
Because the DNA is not denatured by alkaline lysis, AquaPlasmid can yield 10-15 ug DNA/ml of culture.
As low as $0.33/miniprep for 5-10 ug of DNA from 0.5 ml overnight culture, sufficient for most applications.
Improve column-based purification
By simply replacing the alkaline lysis solution (0.2 N NaOH and 1% SDS) in your column-based purification kit with AquaPlasmid solution for cell lysis, you will instantly improve your plasmid kit and at least double its plasmid DNA yield. We can offer AquaPlasmid solution as OEM to interested plasmid kit manufacturers for their column-based purification.
Comparison of AquaPlasmid and column-based plasmid purification
1. Harvest the Cells
Transfer 1.5 ml of overnight bacterial culture to a 1.7-ml microfuge tube. Centrifuge at 14,000 xg for 1 min at room temperature (~22 °C) to pellet the bacteria. Aspirate or flip the tube forcefully a few times to remove the culture medium as completely as possible.
2. Lyse the Cells
Add 100 ul of deionized water to the cell pellet. Vortex at top speed to fully resuspend the cells. Add 100 ul of AquaPlasmid solution to the cell suspension and immediately touch-vortex (a few seconds on and a few seconds off) 5-10 times to mix the contents well. Incubate at room temperature for 5-10 min to fully lyse the cells.
3. Remove the Debris
Add 50 ul of DR solution to the crude lysate. Vortex vigorously at top speed to break up the aggregates into fine pieces. Centrifuge at 14,000 xg for 5 min to pellet the debris.
4. Pellet the Plasmid DNA
Transfer 200 ul of the clear lysate to a clean microfuge tube. Add 0.75 vol (150 ul) of isopropanol to the lysate (Make sure not to use more than 1 vol of isopropanol for DNA precipitation). Touch-vortex at top-speed 5-10 times to mix well. Immediately centrifuge at 14,000 xg at room temperature for 5 min to pellet the plasmid DNA. Flip the tube to discard the supernatant. Fill the tube with 70% ethanol from a squirt bottle (be sure to rinse the inside of the lid as well), and then flip the tube to discard the ethanol solution. Repeat the ethanol rinse once. Flip the tube forcefully 5-6 times and blot it on a paper towel to remove residual ethanol. Place the tube upside down on the paper towel to air-dry the DNA pellet for 5-10 min. Add 100 ul of deionized water to the DNA pellet, vortex to solubilize the DNA.
Product Name: AquaPlasmidTM Kit
Product Number: 1001, 1030
Application: Isolation of plasmid DNA. For in vitro research use only.
Size: The kit is sufficient for the preparation of
Kit Contents: The AquaPlasmid Kit includes the following items
Ordering: To order, please click the "BUY" button below.
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