ZymoPURE™ Plasmid Midiprep Kit

The fastest, easiest, most reliable method for purification of up to 300 µg of ultra-pure endotoxin-free plasmid DNA.
Routinely recover ≥ 1 µg/µl plasmid DNA directly from a microcentrifuge column that is ideal for transfection and other sensitive downstream applications.

Product Size Catalog # Price Qty
ZymoPURE™ Plasmid Midiprep Kit 25 Preps D4200
$255.00
ZymoPURE™ Plasmid Midiprep Kit 50 Preps D4201
$458.00

About ZymoPURE™ Plasmid Midiprep 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 Midiprep kit features a spin column-based method for the purification of up to 300 µg of high-quality plasmid DNA in less than 18 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 Midiprep contains colored buffers that permit error-free visualization and identification of complete bacterial cell lysis and neutralization. Syringe filters are included for rapid clearing of the lysate and the unique spin-column design allows the binding step to be performed using a vacuum or table top centrifuge.

*Patent Pending


Processing Time 18 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 300 µg per preparation (Actual yield is dependent on the plasmid copy number, culture growth conditions, and strain of E. coli utilized)
Recovery Volume ≥ 100 µl of ZymoPURE Elution Buffer or DNase free water
Equipment Needed Microcentrifuge and vacuum/vacuum manifold (recommended) or swinging bucket centrifuge.
Recommended Sample Volume 50 ml 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 8 ml of ZymoPURE™ P1 (Red)to the bacterial cell pellet and resuspend completely by vortexing or pipetting.
  2. Add 8 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 8 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. Ensure the plug is attached to the Luer Lock at the bottom of the ZymoPURE™Syringe Filter. Place the syringe filter upright in a tube rack and load the lysate into the ZymoPURE™Syringe Filter and wait 5 - 8 minutes for the precipitate to float to the top.
  5. Remove the Luer Lock plug from the bottom of the syringe and place it into a clean 50 ml conical tube. Place the plunger in the syringe and push the solution through the ZymoPURE™Syringe Filter to clear the debris. Save the cleared lysate!
  6. Add 8 ml ZymoPURE™Binding Buffer to the cleared lysate from step 6 and mix thoroughly by inverting the capped tube 8 times.
  7. Vacuum Protocol

    The vacuum pump should be a single
    or double-staged unit capable of
    producing up to 400 mm Hg pressure.

    Centrifugation protocol

    A swinging bucket rotor is required for centrifugation.

    8. Ensure the connections of the Zymo-Spin™ III-P Column Assembly are finger-tight and place onto a vacuum manifold.

    9. Add the entire mixture from step 7 into the Zymo-Spin™ III-P Column Assembly, and then turn on the vacuum until all of the liquid has passed completely through the column.

    10. Unscrew the purple Luer Lock cap from the top of the Zymo-Spin™ III-P Column and discard the Reservoirs.

    8. Remove the 50 ml Reservoir from the top of the Zymo-Spin™III-P Column Assembly. Ensure the connection between the 15 ml Conical Reservoir and Zymo- Spin™ III-P colum nis finger-tight and place the assembly into a 50 ml conical tube.

    9. Add 10 ml of the mixture from step 7 into the 15 ml Conical Reservoir/Zymo-Spin™ III-P Column assembly, and centrifuge at 500 x g for 2 minutes.

    10. Empty the 50 ml conical tube and repeat step 9 until the entire sample has passed through the column.

Note: Steps 11-12 can also be completed using a microcentrifuge instead of the vacuum manifold (see full instruction manual).

  1. With the vacuum off, add 800 µl of ZymoPURE™Wash 1 to the Zymo-Spin™III-P Column.Turn on the vacuum until all of the liquid has passed completely through the column.
  2. With the vacuum off, add 800 µl of ZymoPURE™Wash 2to the Zymo-Spin™III-P Column. Turn on the vacuum until all of the liquid has passed completely through the column. Repeat this wash step.
  3. Place the Zymo-Spin™III-P Column in a Collection Tube and transfer to a microcentrifuge. Centrifuge at ≥10,000 x gfor 1 minute in order to remove any residual wash buffer.
  4. Transfer the Zymo-Spin™III-P Column into a clean 1.5 ml tube and add 200 µl of ZymoPURE™Elution Buffer directly to the column matrix. Incubate at room temperature for 2 minutes, and then centrifuge at ≥10,000 x g for 1 minute in a microcentrifuge.
For specific notes and additional information, please see the product protocol PDF.
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“With your kit, I got twice the amount of DNA that I usually get with Invitrogen's midi kit, in half the time. Can't beat that!”
D. G., Immunology Consultants Laboratory
“This kit was absolutely amazing, quicker and easier to use than the Qiagen kit I have used previously
E. S., Queen Mary University of London
“I have used the Promega and Qiagen kits, but even with following all the steps carefully, I have never got such high quality and concentrated DNA.”
P. J., University of Miami
“This kit is superior to the Qiagen kit in speed of preparation, amount of plasmid recovered and fewer (and easier) protocol steps.”
R. K., University of Alabama at Birmingham
“I like this midiprep kit alot!! It can purify the plasmids with high quality in short period of time.”
T. S., National Cancer Centre Singapore

This is what I like about the kit

  1. high purity and yield.
  2. the protocol is short and easy to follow,
  3. there is an option of using of vacuum or centrifuge
  4. coloured buffers
  5. consumables are provided with the kit
M. C., University of Pretoria
“VERY fast protocol, Great color coding of the buffers, nanodrop curves looked perfectly fine, purity and quality perfect for my uses.”
K. G., Friedrich-Alexander University Erlangen-Nuremberg
“My student had an easy time with this kit also, and he had never done a DNA prep before.”
T. W., Northeastern University
“This kit it pretty great, midiprep DNA in about the time it takes to do a miniprep.”
J. G., Yale University
“It went beyond my expectation. The protocol is very easy to follow. Very good DNA yields and the DNA was high quality”
E. P., USDA-Agriculture Research Service
Click here to submit your review of the ZymoPURE™ Plasmid Midiprep Kit.

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