AquaPlasmidTM is a multifunctional reagent for plasmid DNA purification without using columns. This single solution completes cell suspension, cell lysis, plasmid DNA extraction and cell debris removal in a single tube. AquaPlasmid is nontoxic. It does not contain hazardous chemicals, such as guanidine hydrochloride, used in column-based methods. The multifunctionality makes AquaPlasmid the most economic ($0.5/miniprep) and environment-friendly (nontoxic) plasmid purification product on the market. The isolated plasmid DNA is pure and suitable for all downstream applications, including automated DNA sequencing.
AquaPlasmid extracts plasmid DNA without using alkaline lysis and without denaturing the DNA.
AquaPlasmid completes cell suspension, cell lysis, plasmid extraction and debris removal in a single tube.
A small 60-ml AquaPlasmid kit can be used for 300 mini, 30 midi, or 3 maxi preps.
No phenol, no chloroform, and no guanidine salts.
Only $0.50/miniprep, 1/3 of column-based purification kits.
AquaPlasmid purified plasmid DNA is pure (A260/A280 = 1.8-1.9) and suitable for routine molecular biology applications, including automated DNA sequencing.
Comparison of AquaPlasmid and column-based plasmid purification
* Deformities in 20-40% of frogs around the cities? 10-20 times more frogs around major cities are found to have deformed limbs. When buffers containing guanidine salts are dumped into the sewage (hundreds of tons a year!), they could react with Bleach from household and industrial washers. The toxic reaction products could contaminate the water and harm the aquatic lives.
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 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 200 ul of AquaPlasmid solution to the cell pellet. Immediately touch-vortex (a few seconds on and a few seconds off) at top speed for 10-20 sec to fully suspend the cells. Incubate at 22 °C for 10 min to lyse the cells.
3. Remove the Debris
Incubate the crude lysate at -20 °C for 5 min or on ice for 10 min to induce precipitation. Centrifuge at 14,000 xg for 5-10 min at 22 °C to pellet the cellular debris. IMPORTANT: Do not invert or vortex or shake the crude lysate after the lysis incubation or it will cause genomic DNA contamination (see the figure below)!
4. Pellet the Plasmid DNA
Transfer the clear lysate (~180 ul, be careful not to transfer any cellular debris) to a clean 0.5-ml microfuge tube. Add 0.5 vol (90 ul) of isopropanol to the lysate (do not use more than 0.7 vol of isopropanol for DNA precipitation to avoid contamination). Touch-vortex at top-speed 5-10 times to mix well. Centrifuge at 14,000 xg for 5 min at 22 °C to pellet the plasmid DNA. Flip the tube (you may hold 6-10 tubes between your thumb and index finger at the same time) forcefully a few times to discard the supernatant.
5. Rinse the Plasmid DNA
To rinse the DNA pellet (nearly invisible), overflow 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 with a swirling motion to discard the ethanol solution. Flip the tube forcefully a few times and blot it on a paper towel to remove residual ethanol. Leave the tube upside down on the paper towel for 5 min to air-dry the DNA pellet. Add 50 ul of deionized water (or TE buffer) to the DNA pellet and vortex for 30 sec to fully solubilize the DNA. Centrifuge at 14,000 xg for 2-3 min to pellet any insoluble (nearly invisible). Optional: transfer the DNA solution to a new tube.
Product Name: AquaPlasmidTM Kit
Product Number: 1001, 1015, 1030, 1060
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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