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DNA & RNA Purification

GenElute™ HP Select Plasmid Gigaprep Kit


Description | Procedure | Application | Beneficial Features | Order Information | Data

Description: The GenElute™ HP Select Plasmid Gigaprep Kit offers a simple, rapid, and cost-effective method for isolating endotoxin-free plasmid DNA from recombinant E. coli cultures. Up to 15 mg of high copy plasmid DNA with ≤ 0.1 Endotoxin Unit/µg can be recovered from E. coli cultures in 1.2–2.5 liters Luria Broth (LB) medium or 600 ml – 1.2 liters of Terrific Broth (TB) medium in less than 2.0 hours.

  • Sophisticated – utilizes HP Select technology, the premier plasmid purification method
  • Novel – includes patent-pending endotoxin removal system
  • Plentiful – up to 15 mgs of high-quality, endotoxin-free (≤0.1EU/µg) plasmid DNA in 2 hours or less
  • Convenient – vacuum format and no ethanol precipitation required

 

Procedure: An overnight recombinant E. coli culture is harvested by centrifugation and subjected to a modified alkaline-SDS lysis. The lysate is cleared by filtration followed by the addition of a binding solution optimized for endotoxin-free plasmid preparations. The plasmid DNA is then captured onto a silica membrane in the presence of high salts, while endotoxins are prevented from adsorbing to the membrane. Contaminants are removed with three wash steps. Finally, the bound DNA is eluted in endotoxin-free water.

 

Prepare binding column.
 

 

Resuspend overnight culture and lyse cells.
 

 

Neutralize and filter lysate.
 

 

 

Load lysate onto binding column and wash.
 

 

 

Elute plasmid DNA.
 

Application: High-quality, endotoxin-free DNA is ready for immediate use for the most demanding applications including transfection with endotoxin-sensitive cells and gene therapy research.


Beneficial Features
Speed

From harvested bacterial culture to pure plasmid DNA in 2 hours or less

Versatile

Can be used to purify low-, medium- and high-copy plasmid DNA

Yield

Up to 15 mg high-copy plasmid DNA

Convenience

Vacuum format eliminates need for slow gravity flow columns

Economic

Lower cost, higher yields and significantly reduced labor equal substantial savings – in both dollars and time!

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Order Information

Product Code Product Name Purifications
per kit
Technical Manual .pdf
NA0800 GenElute HP Select Plasmid Gigaprep Kit 5 X

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Sample Data

Table 1. Comparison of plasmid yield and time/prep for the different purification systems used for isolating medium-copy number plasmid pFlag-CMV1/DH5 in LB medium.


Purification Kit Plasmid Yield (mg) Time/Prep (hours) Alcohol Precipitation Required?
GenElute HP Select Plasmid Gigaprep Kit 12.23 1.5 NO
Supplier Q Endo-Free Plasmid Giga Kit 10.87 4.8 YES
Supplier B Plasmid Giga EF Kit 6.27 3.3 YES

 

 

Figure 1. Maximum plasmid DNA yield obtained with different constructs using GenElute HP Select Plasmid Gigaprep Kit.


Graph for plasmid DNA yield

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Figure 2. Comparison of transfection efficiencies of high copy plasmid DNA pCMV-SPORT-β-gal/DH5α into endotoxin sensitive cell line HuH-7 cells. The data shows the average and standard deviations of three replicates for the cell line evaluated. Untransfected HuH-7 represent cells treated with transfection reagents without any plasmid DNA.


Graph for plasmid DNA yield

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