ZymoPURE II Plasmid Maxiprep Kit
| Cat# | Name | Size |
|---|---|---|
| D4203 | ZymoPURE II Plasmid Maxiprep Kit | 20 Preps. |
| D4202 | ZymoPURE II Plasmid Maxiprep Kit | 10 Preps |
Documents
Protocol:
Datasheet"
SDS (MSDS):
Description
Highlights
- Fastest: Simple 20 minute Maxipreps.
- Highest Yield: Purify up to 1.2mg 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 Maxiprep kit provides the fastest and simplest method available to efficiently isolate up to 1.2 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 Maxiprep kit can process up to 150 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 Maxiprep 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
1.2 mg
Culture Input
≤ 150 ml
Elution Volume
≤ 0.025 EU/µg Plasmid DNA
Endotoxin Levels
Cells, tissues, blood, plasma, serum, saliva, urine, feces, environmental samples, etc.
Equipment
Microcentrifuge and vacuum/vacuum manifold (recommended) or swinging bucket centrifuge.
Format
Spin Column
Processing Time
20 min
Purity
Typical Abs 260/280 ≥1.8 and Abs 260/230≥ 2.0
Recommended Sample Volume
≤ 150 ml
Size Range
Up to 200 kb
Yield
≤ 1.2 mg per preparation (Actual yield is dependent on the plasmid copy number, culture growth conditions, and strain of E. coli utilized)
Q1: How do I elute from the included Spin-Column?
The design of the Zymo-Spin V-P Column does make the elution step a little tricky. Sometimes air bubbles can develop while adding the elution buffer to the column, which will result in not being able to add all 400 ul to the column. In order to avoid this, we recommend inserting the tip of the micro pipet directly into the column at an angle until the tip is touching the matrix. Then, slowly expel the elution buffer while removing the tip from the column. Some end users find it easier to use the smaller 200 ul pipet tip and load the column twice with 200 ul using the above technique.
Q2: What is the composition of the ZymoPURE Elution buffer?
10 mM Tris-HCl, 0.1 mM EDTA, pH 8.5.
Q3: Can the ZymoPURE Kits be used with other bacteria?
Yes, please contact technical support for an application note.
Q4: 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.
Q5: Are the ZymoPURE kits compatible with any commercially available vacuum manifold?
Yes, any vacuum that uses standard luer-lock connectors is compatible.
Q6: 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.
Q7: 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.
Q8: 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.
Q9: 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 to overloading and subsequent clogging of columns, which can reduce DNA yield/quality or result in failed preps.
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),
“Perfect! Finally, we got 600ug plasmid from 150ml E.coli culture in half an hour.”
– B. P., Los Angeles Biomedical Research Institute at Harbor-UCLA
“We were impressed by the high yield of plasmid DNA from this kit! From a 150 mL culture, I was able to isolate 0.912 mg DNA, in the same amount of time (and effort) I could get about half as much DNA from the comparable Qiagen kit.”
– C. S., Abbott Laboratories
“I was very satisfied with this product. It produced high DNA concentration eluting in 400 µl using the above mentioned cell line, which would generally only be obtainable with other kits eluting in much smaller volumes. This product is also miles ahead of other general Maxiprep kits in terms of time required for plasmid extraction.”
-J. P., University of Pretoria
| Cat # | Name | Size |
|---|---|---|
| D4200-1-150 | ZymoPURE P1 (Red) | 150 ml |
| D4200-2-150 | ZymoPURE P2 (Green) | 150 ml |
| D4200-3-150 | ZymoPURE P3 (Yellow) | 150 ml |
| D4200-4-150 | ZymoPURE Binding Buffer | 150 ml |
| D4200-5-55 | ZymoPURE Wash 1 | 55 ml |
| D4200-6-23 | ZymoPURE Wash 2 (Concentrate) | 23 ml |
| D4200-7-6 | ZymoPURE Elution Buffer | 6 ml |
| C1036-5 | ZymoPURE Syringe Filters | 5 Pack |
| C1037-5 | ZymoPURE Syringe Plungers | 5 Pack |
| C1042-5 | Zymo-Spin V-P Column Assembly w/ 15 ml and 50 ml Reservoir | 5 Pack |
| C1051-10 | EndoZero Spin Columns | 10 Pack |
- Catalog#: D4203 /D4202
- Package Length (in Inches): 13
- Package Width (in Inches): 8.5 /8
- Package Height (in Inches): 13 /6.5
- Package Weight (in Pounds): 10 /4.8
- Size: 20 Preps /10 Preps
- Unit Standard: Metric
- Volume Units: Milliliters
