- Fastest: Simple 20 minute Midipreps.
- Highest Yield: Purify up to 400 µg of plasmid DNA directly from a spin-column.
- Ultra-Pure: EndoZero, vaccine grade, and transfection ready.
Part of the ZymoPURE Plasmid Kits collection, the ZymoPURE II Plasmid Midiprep kit provides the fastest and simplest method available to efficiently isolate up to 400 µg of transfection grade plasmid DNA from E. coli. Utilizing a modified alkaline lysis in conjunction with a novel binding system, the ZymoPURE II Midiprep kit can process up to 50 ml of culture in less than 20 minutes. This remarkable kit results in significantly more plasmid DNA while providing drastically reduced processing times. The plasmid DNA is bound onto a column with either a vacuum or a centrifuge instead of a lengthy gravity flow column. Additionally, there is no ethanol precipitation step required and the elution is performed using a microcentrifuge. The recovered plasmid DNA is highly concentrated (up to 3 µg/µl), EndoZero, vaccine-grade, and transfection-ready. As an added convenience, the ZymoPURE II Plasmid Midiprep kit contains colored buffers that permit error-free visualization and identification of complete bacterial cell lysis and neutralization.
|Applicable For||Transfection, transformation, sequencing, restriction endonuclease digestion, in vitro transcription, in vivo studies, genome editing, and other sensitive applications.|
|Binding Capacity||400 µg|
|Culture Input||≤ 50 ml|
|Elution Volume||≥ 100 µl|
|Endotoxin Levels||≤ 0.025 EU/µg Plasmid DNA|
|Equipment||Microcentrifuge and vacuum/vacuum manifold (recommended) or swinging bucket centrifuge|
|Processing Time||20 min|
|Purity||Typical Abs 260/280 ≥1.8 and Abs 260/230≥ 2.0|
|Size Range||Up to 200 kb|
|Yield||≤ 400 µg per preparation (Actual yield is dependent on the plasmid copy number, culture growth conditions, and strain of E. coli utilized)|
Q1: What is the composition of the ZymoPURE Elution buffer?
10 mM Tris-HCl, 0.1 mM EDTA, pH 8.5.
Q2: Can the ZymoPURE Kits be used with other bacteria?
Yes, please contact technical support for an application note.
Q3: What type of vacuum pump do you recommend?
The vacuum pump should be a single or double-staged unit capable of producing at least 400 mm Hg pressure at the vacuum manifold. If less pressure is applied, centrifuge the column prior to washing to remove any residual lysate/buffer remaining in the matrix.
Q4: Are the ZymoPURE kits compatible with any commercially available vacuum manifold?
Yes, any vacuum that uses standard luer-lock connectors is compatible.
Q5: Can an in-house vacuum line be used with the EZ Vac Vacuum Manifold?
Yes, however, the pressure needs to be around 400 mm Hg. Users should take caution, as the pressure of an in-house vacuum line can fluctuate drastically or be significantly reduced, depending on the demand in the building.
Q6: I ran out of ZymoPURE Wash 2. Can I substitute it with a homemade solution or Wash Buffer from another kit?
No, the ZymoPURE kits are only compatible with ZymoPURE Wash 2. Additional ZymoPURE Wash 2 can be purchased separately.
Q7: Is there a protocol for low-copy number plasmid DNA?
Yes, the low-copy protocol can be found in Appendix A of the kit protocol.
Q8: For low-copy plasmids, can I exceed the recommended volume of bacterial culture or use enriched growth media?
Yes, however, special care should be taken throughout the entire protocol since there is much more biomass. We recommend centrifuging the neutralized lysate before loading onto the supplied syringe filter. For further guidance on dealing with low-copy plasmids, please contact our technical support team. Exceeding the recommended volume for high copy plasmids can cause to overloading and subsequent clogging of columns, which can reduce DNA yield/quality or result in failed preps.
Q9: Can the ZymoPURE kits be used to isolate large plasmid constructs (BAC/PAC)?
Yes, the standard ZymoPURE protocol has been successfully tested with constructs up to 200 kb. To increase elution of large plasmid DNA, we recommend pre-warming the ZymoPURE Elution Buffer (50 ºC) and increasing the incubation time on column up to 10 minutes prior to centrifugation.
“It went beyond my expectation. The protocol is very easy to follow. Very good DNA yields and the DNA was high quality”
- J. G., Yale University
“This kit it pretty great, midiprep DNA in about the time it takes to do a miniprep.”
- T. W., Northeastern University
“My student had an easy time with this kit also, and he had never done a DNA prep before.”
- K. G., Friedrich-Alexander University Erlangen-Nuremberg