Yields can reach as high as 350 mg for a midiprep and 1.2 mg for a maxiprep. As with any protocol, lower yields can be expected with low copy plasmids.
NA0410
GenElute™ HP Endotoxin-Free Plasmid Maxiprep Kit
sufficient for 25 preparations
Synonym(s):
Gen Elute, GenElute Endotoxin-Free Plasmid Kit
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Product Name
GenElute™ HP Endotoxin-Free Plasmid Maxiprep Kit, sufficient for 25 preparations
usage
sufficient for 25 preparations
greener alternative product characteristics
Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.
sustainability
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technique(s)
DNA extraction: suitable
Quality Level
test parameters
sample volume: 150 mL bacterial culture
greener alternative category
storage temp.
15-25°C
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This Item | NA0310 | PLEX15 | PLX15 |
|---|---|---|---|
| technique(s) DNA extraction: suitable | technique(s) DNA purification: suitable | technique(s) DNA purification: suitable | technique(s) DNA purification: suitable |
| usage sufficient for 25 preparations | usage sufficient for 25 purifications | usage sufficient for 15 purifications | usage sufficient for 15 purifications |
| greener alternative product characteristics Designing Safer Chemicals | greener alternative product characteristics Designing Safer Chemicals | greener alternative product characteristics Inherently Safer Chemistry for Accident Prevention | greener alternative product characteristics Designing Safer Chemicals |
| sustainability Greener Alternative Product | sustainability Greener Alternative Product | sustainability Greener Alternative Product | sustainability Greener Alternative Product |
NA0400 | NA0300, NA0300S | - | - |
| greener alternative category | greener alternative category | greener alternative category , Aligned | greener alternative category |
Application
Biochem/physiol Actions
An overnight recombinant E. coli culture is harvested by centrifugation and subjected to a modified alkaline-SDS lysis. The lysate is clarified by filtration followed by the addition of a binding solution that has been optimized for endotoxin-free plasmid preparations. The plasmid DNA is then captured on silica, while endotoxins are prevented from adsorbing to the membrane. Two wash steps remove contaminants. Finally, the bound DNA is eluted in endotoxin-free water. The recovered plasmid DNA is predominately in its supercoiled form. Genomic DNA and RNA are below detectable levels by ethidium bromide stained agarose gel electrophoresis.
Features and Benefits
- Up to 1.2 mg of high-copy plasmid DNA with endotoxin levels of ≤0.1 EU/μg
- Only 35 minutes from pelleted cells to purified plasmid
- Convenient vacuum format
General description
The GenElute HP Endotoxin-Free Plasmid Maxiprep Kit offers a simple and rapid method for isolating endotoxin-free plasmid DNA from recombinant E. coli cultures. The kit uses a vacuum format with a filter column for the rapid clearing of the bacterial lysate and a silica column for capturing plasmid DNA. A proprietary solution binds plasmid DNA to the binding column while preventing endotoxins from adsorbing. The technology allows for the user to consistently achieve levels of endotoxin that are less than 0.1 endotoxin units per μg of plasmid DNA.
High-quality, endotoxin-free DNA is ready for immediate use for the most demanding applications including transfection with endotoxin-sensitive cells.
High-quality, endotoxin-free DNA is ready for immediate use for the most demanding applications including transfection with endotoxin-sensitive cells.
Other Notes
For additional information, please see www.sigma-aldrich.com/genelutehp.
Legal Information
GenElute is a trademark of Sigma-Aldrich Co. LLC
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Product No.
Description
Pricing
signalword
Danger
Hazard Classifications
Acute Tox. 4 Oral - Eye Irrit. 2 - Flam. Liq. 3 - Met. Corr. 1 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
target_organs
Central nervous system
Storage Class
3 - Flammable liquids
flash_point_f
77.0 °F - closed cup
flash_point_c
25 °C - closed cup
wgk
WGK 3
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Related Content
GenElute HP Plasmid Maxiprep Kit isolates endotoxin-free plasmid DNA in 35 minutes, meeting industry standards.
Tools for plasmid preparation, in vitro transcription, mRNA purification, and LNP formulation in mRNA-based vaccine and therapeutic development.
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What is the expected yield of plasmid DNA that you can isolate from these kits?
1 answer-
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How long does it take to isolate plasmid using the vacuum format?
1 answer-
A midiprep or maxiprep requires only 30 minutes to complete.
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How do the GenElute HP Plasmid Kits differ from other plasmid purification systems?
1 answer-
The GenElute HP Plasmid Kits are much faster than other commercially available kits. A filter syringe is used for preparing the cleared lysate, which replaces a lengthy centrifugation step. These kits also offer the option of using a vacuum or spin purification format rather than a gravity drip to bind, wash and elute the plasmid. In addition, samples are eluted in Tris buffer so there are no alcohol precipitations required.
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Do the binding columns fit on other manufacturers’ manifolds?
1 answer-
Yes. The columns fit on all standard vacuum manifolds.
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Which format gives a better yield, vacuum or spin?
1 answer-
There is no difference in plasmid recovery when using either format.
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Is plasmid DNA isolated with this kit suitable for use in transfection?
1 answer-
Yes. However, if you are currently required to use an endofree plasmid purification kit, we recommend using the GenElute HP Endotoxin-free Kit.
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