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Nucleic Acid Purification


Products >  Nucleic_Acid_Purification >  Plasmid >  Endotoxin_Free >  Preparative_Scale

Preparative Scale Plasmid DNA Purification, Endotoxin-Free—NucleoBond PC Prep 100

Endotoxins (lipopolysaccharides, or LPS) are a major component of the Gram-negative bacterial cell wall. The LPS molecule is an extremely potent stimulator of the mammalian immune system, and a number of mechanisms exist to detect LPS and to respond to the presence of either this molecule or Gram-negative bacteria. LPS is a common contaminant of plasmid DNA preparations grown in E. coli. The negative charges associated with lipid A and the inner core of LPS cause the LPS molecule to behave like DNA on anion exchange chromatography resins. An important prerequisite for transfections and gene therapy applications is an endotoxin-free plasmid DNA.

Minimizing the level of endotoxin contamination in purified plasmid DNA is critical for many applications. For this reason MACHEREY-NAGEL developed a specific patented procedure that reduces endotoxins to a very low level. Unlike competitor kits, there is no need for a time-consuming incubation step to remove the endotoxins (see procedure). The working procedure for the endotoxin-free plasmid preparation is identical to the standard protocol, and each prep yields up to 100 mg of endotoxin-free plasmid DNA.

NucleoBond EF kits employ a modified alkaline/SDS lysis procedure to prepare the bacterial cell pellet for plasmid purification. The bacterial lysate is cleared by filtration and loaded onto the equilibrated column. No additional incubation step is required for endotoxin removal. After efficient washing of the column, the purified plasmid DNA is eluted from the column.

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Features

  • Endotoxins are efficiently removed (<0.1 EU/µg DNA)
  • No additional incubation is required for endotoxin removal
  • Purify up to 100 mg from 5–20 L of overnight E. coli culture
  • Patented procedure

Applications

Purified DNA is suitable for:

  • Transfection of highly sensitive cells
  • Lentivirus production

Product Overview

Technology Anion exchange
Format Preparative scale
Lysate Clarification Folded filters
Starting Material 5–20 L
Vector Size <300 kb
Typical Yield 80–100 mg
A260/280 1.80–1.95
Endotoxin Level <0.1 EU/µg
Preparation Time 20 hr/prep

 
 
Products
Cat. # Product Contents Size Price Units Select
740594 NucleoBond® PC Prep100 Each $1,650.00
 

 

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