ZymoPURE - Express Plasmid Midiprep Kit
ZymoPURE - Express Plasmid Midiprep Kit
- Direct lysis protocol omits conventional cell-pelleting and resuspension steps.
- Purify up to 1.2 mg of highly concentrated plasmid DNA directly from a spin-column.
- Prepare transfection-grade plasmid DNA in only 15 minutes.
Part of the ZymoPURE Plasmid Kits collection, the ZymoPURE – Express Plasmid Midiprep kit utilizes a patented alkaline lysis method for purifying up to 1.2 mg of transfection-grade plasmid DNA directly from E. coli culture without conventional cell-pelleting and resuspension steps. The ZP Express Lysis Buffer is simply added directly to the culture and the plasmid DNA is purified following neutralization and removal of debris using the patented ZymoPURE binding system.
The plasmid DNA is rapidly bound onto a column with a vacuum or a centrifuge instead of a slow gravity flow column. Additionally, there is no alcohol precipitation step required and the elution is performed using a microcentrifuge. The ZymoPURE – Express Plasmid Midiprep kit’s fast and simple protocol enables the recovery of highly concentrated transfection-grade plasmid DNA in just 15 minutes. As an added convenience, the ZymoPURE – Express Plasmid Midiprep kit contains colored buffers that permit error-free visualization and identification of complete bacterial cell lysis and neutralization.Learn More
|Applicable For||Transfection, transformation, lentivirus production, adenovirus production, AAV production, CRISPR, genome editing, sequencing, restriction endonuclease digestion, in vitro transcription/translation, PCR, and other sensitive applications.|
|Binding Capacity||1.2 mg|
|Culture Input||25 ml|
|Elution Volume||≥ 150 µl|
|Endotoxin Levels||≤1 EU/µg of Plasmid DNA|
|Equipment||Microcentrifuge and vacuum/vacuum manifold (recommended) or swinging bucket centrifuge|
|Processing Time||15 min|
|Purity||Typical Abs260/280 ≥1.8 and Abs260/230≥ 2.0|
|Recommended Sample Volume||≤ 25 ml|
|Size Range||Up to 200 kb|
|Yield||Up to 1.2 mg per preparation. Actual yield is dependent on the plasmid copy number, culture growth conditions, and strain of E. coli utilized. Typical yields from 25 ml of overnight culture grown in LB for high copy number plasmids are 100 – 200 µg.|
Q1: What is the composition of the ZymoPURE Elution buffer?
10 mM Tris-HCl, 0.1 mM EDTA, pH 8.5.
Q2: I accidently left my ZP Express Neutralization Buffer at room temperature, Is it still okay to use?
Yes, the RNase A is fairly stable at room temperature but we recommend placing it in 4 °C as soon as possible to ensure optimal performance throughout the life span of the product.
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 for the liquid to pass through the columns quickly. 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 and we cannot disclose a substitution due to the sensitive nature of the recipe. Additional ZymoPURE Wash 2 can be purchased separately.
Q7: Can you recover up to 1.2 mg of plasmid with the ZymoPURE-Express Plasmid Midiprep Kit?
The binding capacity of the column is up 1.2 mg of plasmid, however, typical plasmid yields from directly processing 25 ml of overnight culture for a high copy plasmid is 100 – 200 µg.
Q8: Can I process more than 25 ml of culture?
Yes, up to 50 ml of bacterial culture grown in LB can be pelleted, resuspended, and processed with the kit. Please see the appendix in the protocol for more information.
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 the elution efficiency of large plasmid DNA, we recommend pre-warming the ZymoPURE Elution Buffer (50 ºC) and increasing the incubation time on the column up to 10 minutes prior to centrifugation.
|D4213-1-210||ZP Express Lysis Buffer||210 ml||$267.30|
|D4213-2-260||ZP Express Neutralization Buffer||260 ml||$82.50|
|D4213-3-260||ZP Express Binding Buffer||260 ml||$79.20|
|D4200-5-20||ZymoPURE Wash 1||20 ml||$18.70|
|D4200-5-55||ZymoPURE Wash 1||55 ml||$44.00|
|D4200-5-320||ZymoPURE Wash 1||320 ml||$214.50|
|D4200-5-410||ZymoPURE Wash 1||410 ml||$286.00|
|D4200-6-10||ZymoPURE Wash 2 (Concentrate)||10 ml||$19.80|
|D4200-6-12||ZymoPURE Wash 2 (Concentrate)||12 ml||$41.80|
|D4200-6-23||ZymoPURE Wash 2 (Concentrate)||23 ml||$44.00|
|D4200-6-28||ZymoPURE Wash 2 (Concentrate)||28 ml||$47.30|
|D4200-7-1||ZymoPURE Elution Buffer||1ml||$11.00|
|D4200-7-6||ZymoPURE Elution Buffer||6 ml||$12.10|
|D4200-7-12||ZymoPURE Elution Buffer||12 ml||$22.00|
|D4200-7-30||ZymoPURE Elution Buffer||30 ml||$41.80|
|C1001-50||Collection Tubes||50 Pack||$16.50|
|C1001-500||Collection Tubes||500 Pack||$57.20|
|C1001-1000||Collection Tubes||1000 Pack||$99.00|
|C1092-5||ZymoPURE Syringe Filter-X||5 Pack||$30.80|
|C1037-5||ZymoPURE Syringe Plungers||5 Pack||$15.40|
|C1083-5||Zymo-Spin V-PS Column Assembly w/15 ml Reservoir-X and 50 ml Reservoir||5 Pack||$72.60|