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NA0410

GenElute™ HP Endotoxin-Free Plasmid Maxiprep Kit

sufficient for 25 preparations

Sustainability features

Synonym(s):

Gen Elute, GenElute Endotoxin-Free Plasmid Kit

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1 kit
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$782.00

About This Item

NACRES:
NA.52
UNSPSC Code:
41105501

$782.00


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usage

sufficient for 25 preparations

Quality Segment

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technique(s)

DNA extraction: suitable

test parameters

sample volume: 150 mL bacterial culture

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,

storage temp.

15-25°C

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 and from competent Escherichia coli cells.

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

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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description

sufficient for 25 preparations

description

sufficient for 10 preparations

description

sufficient for 15 purifications

description

sufficient for 25 purifications

usage

sufficient for 25 preparations

usage

sufficient for 10 preparations

usage

sufficient for 15 purifications

usage

sufficient for 25 purifications

technique(s)

DNA extraction: suitable

technique(s)

DNA extraction: suitable

technique(s)

DNA purification: suitable

technique(s)

DNA purification: suitable

Quality Level

200

Quality Level

200

Quality Level

200

Quality Level

200

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Kit Components Also Available Separately

Product No.
Description
SDS & Pricing

  • Red Vac Cap 5
    SDS

  • Column Preparation Solution 2
    SDS

  • Wash Solution 1
    SDS

  • Neutralization Solution EF
    SDS

  • Wash Solution 2
    SDS

  • E-Toxate™ Water, endotoxin, free
    SDS

  • Lysis Solution
    SDS

See All (10)

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

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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Questions

1–6 of 6 Questions  
  1. Do the binding columns fit on other manufacturers’ manifolds?

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

      Helpful?

  2. 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?

  3. Which format gives a better yield, vacuum or spin?

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

      Helpful?

  4. 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?

  5. 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?

  6. 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?

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