ZymoPURE™ Plasmid Gigaprep Kit

The fastest, easiest, most reliable method for purification of up to 10 mg of ultra-pure endotoxin-free plasmid DNA.
Routinely recover ≥ 1 µg/µl plasmid DNA directly from a spin-column that is ideal for transfection and other sensitive downstream applications.

Product Size Catalog # Price Qty
ZymoPURE™ Plasmid Gigaprep Kit 5 Preps D4204
$462.00

About ZymoPURE™ Plasmid Gigaprep Kit

ZymoPURE Plasmid Isolation kits provide the fastest and simplest method available to efficiently isolate transfection quality plasmid DNA from E. coli. The ZymoPURE Plasmid Gigaprep kit features a spin column-based method for the purification of up to 10 mg of high-quality plasmid DNA in less than 50 minutes. The eluted plasmid DNA is ready for immediate use, avoiding the need for subsequent precipitation steps.

ZymoPURE technology uses a modified alkaline lysis method and features novel binding chemistry that yields highly concentrated plasmid DNA (up to 3 µg/µl) directly from a spin column. The wash regimen has been optimized to ensure the plasmid DNA is free of endotoxins, salt, protein, and RNA resulting in plasmid DNA suitable for sensitive applications including transfection, plasmid-mediated gene silencing, cloning, and sequencing.

As an added convenience, the ZymoPURE Plasmid Gigaprep contains colored buffers that permit error-free visualization and identification of complete bacterial cell lysis and neutralization. Also, bottle top filters are included for rapid clearing of the lysate.

*Patent Pending


Processing Time 50 min
DNA Purity Eluted DNA is ultrapure and well suited for transfection, sequencing, cloning, plasmid mediated gene silencing, in vitro transcription, and other sensitive applications.
Plasmid DNA Size Up to 25 kb
Plasmid DNA Yield Up to 10 mg per preparation (Actual yield is dependent on the plasmid copy number, culture growth conditions, and strain of E. coli utilized)
Recovery Volume ≥ 2 ml of ZymoPURE Elution Buffer or DNase free water
Equipment Needed Vacuum/vacuum manifold and swinging bucket centrifuge.
Recommended Sample Volume 2.5 L E. coli Culture

Yield, concentration and transfection efficiency for plasmid DNA isolated using the ZymoPURE Maxiprep kit compared to two separate kits from Supplier Q. Plasmid DNA (pGL3®) was isolated from 150 ml of JM109 E. coli culture grown overnight following the manufacturer’s suggested protocol (in duplicate). HeLa cells cultured in a 96-well plate were transfected with 100 ng of pGL3® Luciferase Reporter Vector using Lipofectamine® 2000 and luciferase expression was measured 48 hours later with the ONE-Glo Luciferase Assay System and Veritas Microplate Luminometer. Shown are means ± SEM of 8 transfections.

 

Plasmid DNA yield and concentration from the ZymoPure Maxiprep kit compared to other major suppliers. Plasmid DNA (pGEM®) was isolated from 150 ml of JM109 E. coli culture grown overnight following the manufacturer’s suggested protocol (in duplicate). One (1) µl of eluted plasmid DNA was visualized post agarose gel electrophoresis. M, ZR 1 kb DNA Marker (Zymo Research).

  1. Add 150 ml of ZymoPURE™ P1 (Red) to the bacterial cell pellet and resuspend completely by vortexing or pipetting.
  2. Add 150 ml of ZymoPURE™ P2 (Green) and immediately mix by gently inverting the tube 6 times. Do not vortex! Let sit at room temperature for 2-3 minutes. Cells are completely lysed when the solution appears clear, purple, and viscous.
  3. Add 150 ml of ZymoPURE™ P3 (Yellow) and mix gently but thoroughly by inversion. Do not vortex! The sample will turn yellow when the neutralization is complete and a yellowish precipitate will form.
  4. Place the ZymoPURE™ Giga Filter onto a 33 mm or 45 mm-neck glass bottle and load the lysate into the ZymoPURE™ Giga Filter. Ensure the ZymoPURE™ Giga Filter is resting securely on top of the glass bottle and wait 10 minutes for the precipitate to float to the top.
  5. Connect the ZymoPURE™ Giga Filter to a vacuum source and turn on the vacuum until all of the liquid has passed completely through the filter. Save the cleared lysate!
  6. Add 150 ml ZymoPURE™ Binding Buffer to the cleared lysate from step 6 and mix thoroughly by inverting the capped bottle 10 times.
  7. Securely attach the 600 ml Reservoir to the top of the Zymo-Spin™ VI-P Column and place onto a vacuum manifold.
  8. Add the entire mixture from step 7 into the 600 ml Reservoir Zymo-Spin™ VI-P Column Assembly, and then turn on the vacuum until all of the liquid has passed completely through the column.
  9. With the vacuum off, add 80 ml of ZymoPURE™ Wash 1 to the 600 ml Reservoir. Turn on the vacuum until all of the liquid has passed completely through the column.
  10. With the vacuum off, add 50 ml of ZymoPURE™ Wash 2to the 600 ml Reservoir. Turn on the vacuum until all of the liquid has passed completely through the column. Repeat this wash step.
  11. Remove and discard the 600 ml Reservoir and place the Zymo-Spin™ VI-P Column in a 50 ml conical tube. Centrifuge at ≥3,400 x g for 10 minutes in order to remove any residual wash buffer.
  12. Transfer the column into a clean 50 ml conical tube and add 3 ml of ZymoPURE™ Elution Buffer directly to the column matrix. Incubate at room temperature for 5 minutes, and then centrifuge at ≥3,400 x g for 5 minutes.
For specific notes and additional information, please see the product protocol PDF.
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