ZymoPURE II Plasmid Gigaprep Kit
| Cat# | Name | Size |
|---|---|---|
| D4204 | ZymoPURE II Plasmid Gigaprep Kit | 5 Preps. |
Documents
Protocol:
Datasheet"
SDS (MSDS):
Description
Highlights
- Fast: 50 minute Gigapreps
- Highest Yield: Purify up to 10 mg of plasmid DNA directly from a spin-column.
- Ultra-Pure: EndoZero, vaccine grade, and transfection ready.
Description
Part of the ZymoPURE Plasmid Kits collection, the ZymoPURE II Plasmid Gigaprep kit provides the fastest and simplest method available to efficiently isolate up to 10 mg of transfection grade plasmid DNA from E. coli. Utilizing a modified alkaline lysis in conjunction with a novel binding system, the ZymoPURE II Plasmid Gigaprep kit can process up to 2.5 L of culture in less than 50 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 a vacuum 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 Gigaprep 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
10 mg
Culture Input
≤ 2.5 L
Elution Volume
≥ 2 ml
Endotoxin Levels
≤ 0.025 EU/µg Plasmid DNA
Equipment
Vacuum/vacuum manifold and swinging bucket centrifuge
Format
Spin Column
Processing Time
50 min
Purity
Typical Abs 260/280 ≥1.8 and Abs 260/230 ≥ 2.0
Recommended Sample Volume
≤ 2.5 L
Size Range
Up to 200 kb
Yield
≤ 10 mg per preparation (Actual yield is dependent on the plasmid copy number, culture growth conditions, and strain of E. coli utilized)
Q1: Can I process more than 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 overloading and subsequent clogging of columns, which can reduce DNA yield/quality or result in failed preps.
Q2: Is there a protocol for low-copy number plasmid DNA?
Yes, please contact our technical support team for a protocol
Q3: 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.
Q4: 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.
Q5: Are the ZymoPURE™ kits compatible with any commercially available vacuum manifold?
Yes, any vacuum that uses standard luer-lock connectors is compatible.
Q6: 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.
Q7: Can the ZymoPURE™ Kits be used with other bacteria?
Yes, please contact technical support for an application note.
Q8: I ran out of ZymoPURE™ Wash 2. Can I substitute
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.
Q9: What is the composition of the ZymoPURE™ Elution buffer?
10 mM Tris-HCl, 0.1 mM EDTA, pH 8.5.
Q10: 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.
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96 / 100 Bioz Stars |
Hectd3 promotes pathogenic Th17 lineage through Stat3 activation and Mal…
Nature Communications – Published 16 Feb 2019
Vector constructs were prepared using ZymoPURE plasmid midiprep kit (CatD4201, Zymo Research, CA, USA) following manufacturer’s instructions.
Molecular safeguarding of CRISPR gene drive experiments
Plasmids used for injection into eggs were purified with ZymoPure Midiprep kit (Zymo Research).
Improving cytidine and adenine base editors by expression optimization a…
Nat Biotechnol – Published 29 Nov 2018
All plasmids were assembled by either the USER cloning method as previously described or by Gibson assembly .. Plasmids for mammalian cell transfections were prepared using an endotoxin removal plasmid purification system, ZymoPURE Plasmid Midiprep (Zymo Research Corporation).. HEK293T cells (ATCC CRL-3216) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Corning) supplemented with 10% fetal bovine serum (FBS),
- Catalog#: D4204
- Package Length (in Inches): 12
- Package Width (in Inches): 10.5
- Package Height (in Inches): 16
- Package Weight (in Pounds): 15.5
- Size: 5 Preps
- Unit Standard: Metric
- Volume Units: Milliliters
