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NA0410

Sigma-Aldrich

GenElute HP Endotoxin-Free Plasmid Maxiprep Kit

greener alternative

sufficient for 25 preparations

Synonym(s):

Gen Elute, GenElute Endotoxin-Free Plasmid Kit

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1 KIT
€677.00

€677.00


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1 KIT
€677.00

About This Item

UNSPSC Code:
41105501
NACRES:
NA.52

€677.00


Please contact Customer Service for Availability

Request a Bulk Order

usage

sufficient for 25 preparations

greener alternative product characteristics

Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.

sustainability

Greener Alternative Product

technique(s)

DNA extraction: suitable

greener alternative category

storage temp.

15-25°C

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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.

Application

GenElute HP Endotoxin-Free Plasmid Maxiprep Kit has been used in the isolation of plasmid from swine liver[1] and from competent Escherichia coli cells.[2]

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

Principle

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.

Other Notes

For additional information, please see www.sigma-aldrich.com/genelutehp.

Legal Information

GenElute is a trademark of Sigma-Aldrich Co. LLC

Signal Word

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 Code

3 - Flammable liquids

Flash Point(F)

77.0 °F - closed cup

Flash Point(C)

25 °C - closed cup


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Towards a metalloprotease-DNA vaccine against piscine cryptobiosis caused by Cryptobia salmositica
Tan CW, et al.
Parasitology Research, 102(2), 265-275 (2008)
Successful liver-directed gene delivery by ERCP-guided hydrodynamic injection (with videos)
Kumbhari V, et al.
Gastrointestinal Endoscopy, 88(4), 755-763 (2018)
Tri-Hung Nguyen et al.
International journal of oncology, 41(3), 829-838 (2012-06-14)
Overexpression of TMPRSS4, a cell surface-associated transmembrane serine protease, has been reported in pancreatic, colorectal and thyroid cancers, and has been implicated in tumor cell migration and metastasis. Few reports have investigated both TMPRSS4 gene expression levels and the protein
Marco Cassano et al.
Development (Cambridge, England), 138(20), 4523-4533 (2011-09-10)
Mice deficient in α-sarcoglycan (Sgca-null mice) develop progressive muscular dystrophy and serve as a model for human limb girdle muscular dystrophy type 2D. Sgca-null mice suffer a more severe myopathy than that of mdx mice, the model for Duchenne muscular
Paul A Beare et al.
Journal of bacteriology, 191(5), 1369-1381 (2008-12-31)
Coxiella burnetii is a gram-negative obligate intracellular bacterium and the causative agent of human Q fever. The lack of methods to genetically manipulate C. burnetii significantly impedes the study of this organism. We describe here the cloning and characterization of

Related Content

GenElute HP Plasmid Maxiprep Kit isolates endotoxin-free plasmid DNA in 35 minutes, meeting industry standards.

Questions

1–6 of 6 Questions  
  1. Which format gives a better yield, vacuum or spin?

    1 answer
    1. There is no difference in plasmid recovery when using either format.

      Helpful?

  2. What is the expected yield of plasmid DNA that you can isolate from these kits?

    1 answer
    1. 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.

      Helpful?

  3. Do the binding columns fit on other manufacturers’ manifolds?

    1 answer
    1. Yes. The columns fit on all standard vacuum manifolds.

      Helpful?

  4. How do the GenElute HP Plasmid Kits differ from other plasmid purification systems?

    1 answer
    1. 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.

      Helpful?

  5. Is plasmid DNA isolated with this kit suitable for use in transfection?

    1 answer
    1. Yes. However, if you are currently required to use an endofree plasmid purification kit, we recommend using the GenElute HP Endotoxin-free Kit.

      Helpful?

  6. How long does it take to isolate plasmid using the vacuum format?

    1 answer
    1. A midiprep or maxiprep requires only 30 minutes to complete.

      Helpful?

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