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ZymoPURE Plasmid Miniprep Kit
D4212 / D4211 / D4210 / D4208T / D4209
- Highest Yield: Purify up to 100 µg of plasmid DNA in as little as 25 µl directly from a spin-column.
- Transfection-Grade: 50,000 times fewer endotoxins than industry leading minipreps.
- BAC/YAC/PAC Ready: Purify DNA up to ~200 kb.
Part of the ZymoPURE™ Plasmid Kits collection, the ZymoPURE™ Plasmid Miniprep Kit features a spin column-based method for the purification of up to 100 µg of ultra-pure endotoxin-free plasmid DNA in less than 15 minutes. The unique spin-column design also provides zero buffer retention and a low elution volume. ZymoPURE™ technology uses a modified alkaline lysis method and features novel binding chemistry that yields highly concentrated plasmid DNA (up to 3 µg/µl). In addition, the wash regimen has been optimized to ensure the plasmid DNA is free of endotoxins, salt, protein, and RNA. The result is plasmid DNA suitable for transfection, restriction endonuclease digestion, bacterial transformation, PCR amplification, DNA sequencing, and other sensitive downstream applications. As an added convenience, the ZymoPURE™ Plasmid Miniprep Kit contains colored buffers that permit error-free visualization and identification of complete bacterial cell lysis and neutralization.
|Applicable For||Transfection, transformation, lentivirus production, adenovirus production, AAV production, CRISPR, genome editing, in vivo studies, sequencing, restriction endonuclease digestion, in vitro transcription/translation, PCR, and other sensitive applications.|
|Binding Capacity||100 µg|
|Culture Input||≤ 5 mL|
|Elution Volume||25 µl|
|Endotoxin Levels||≤ 1 EU/µg|
|Equipment||Microcentrifuge and/or vacuum manifold|
|Processing Time||15 min|
|Purity||Typical Abs 260/280 ≥1.8 and Abs 260/230≥ 2.0|
|Recommended Sample Volume||≤ 5.0 ml|
|Size Range||Up to ~200 kb|
|Yield||Up to 100 µg per preparation, depending 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: 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.
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: Can the ZymoPURE Kits be used with other bacteria?
Yes, please contact technical support for an application note.
Q7: Is there a protocol for low-copy number plasmid DNA?
Yes, the low-copy protocol can be found in the Appendix of the kit protocol.
Q8: Can I process more than the recommended volume of bacterial culture or use enriched growth media?
Yes, however, more attention is required during the lysis and steps. We recommend centrifuging/pelleting the neutralized lysate before loading onto the supplied syringe/bottle-top filter. For high copy plasmids using more culture can cause overloading and subsequent clogging of columns, which can result in poor DNA yield and/or quality.
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.
Q10: Can the ZymoPURE Kits be used with other bacteria?
Yes, the standard protocol should work with other gram-negative bacteria. However, the user will need to validate the protocol for their particular species. For gram-positive bacteria, please refer to the Gram-Positive Bacteria Protocol in the Appendix of the kit protocol.
|C1086-50||Zymo-Spin II-PX Columns||50 Pack|
|C1001-50||Collection Tubes||50 Pack|
|D4200-6-23||ZymoPURE Wash 2 (Concentrate)||23 ml|
|D4200-4-3||ZymoPURE Binding Buffer||3 ml|
|D4200-5-20||ZymoPURE Wash 1||20 ml|
|D4200-5-320||ZymoPURE Wash 1||320 ml|
|D4200-7-6||ZymoPURE Elution Buffer||6 ml|
|D4200-6-28||ZymoPURE Wash 2 (Concentrate)||28 ml|
|D4200-7-12||ZymoPURE Elution Buffer||12 ml|
|D4200-3-210||ZymoPURE P3 (Yellow)||210 ml|
|D4200-3-3||ZymoPURE P3 (Yellow)||3 ml|
|D4200-4-110||ZymoPURE Binding Buffer||110 ml|
|D4200-4-14||ZymoPURE Binding Buffer||14 ml|
|D4200-2-210||ZymoPURE P2 (Blue)||210 ml|
|D4200-2-3||ZymoPURE P2 (Blue)||3 ml|
|D4200-3-100||ZymoPURE P3 (Yellow)||100 ml|
|D4200-3-13||ZymoPURE P3 (Yellow)||13 ml|
|D4200-1-210||ZymoPURE P1 (Red)||210 ml|
|D4200-1-3||ZymoPURE P1 (Red)||3 ml|
|D4200-2-100||ZymoPURE P2 (Blue)||100 ml|
|D4200-2-13||ZymoPURE P2 (Blue)||13 ml|
|D4200-1-100||ZymoPURE P1 (Red)||100 ml|
|D4200-1-13||ZymoPURE P1 (Red)||13 ml|
|D4200-7-30||ZymoPURE Elution Buffer||30 ml|