Obtain High Yields of Transfection-Grade Plasmid DNA—Fast!

• Very pure DNA ensures high transfection efficiency
• Rapid, simple protocol
• Specialized “EF“ kits provide endotoxin-free DNA for use in sensitive cells

Ensure Higher Transfection Efficiency

Transfection efficiency depends on many factors, including DNA quality and purity. Anion-exchange columns (such as NucleoBond Xtra) provide the purest DNA and the best transfection results. However, DNA purified with other brands of anion-exchange DNA purification columns cannot match the purity and transfection efficiency of DNA purified with NucleoBond Xtra DNA purification columns.

Figure 1. Plasmid DNA for a gephyrin-GFP fusion was purified with NucleoBond Xtra and with a plasmid DNA purification kit from Competitor Q. HEK 293 cells were transfected with plasmid DNA and incubated for 48 hr before visualization by fluorescent microscopy. NucleoBond Xtra gave higher transfection efficiency.

Purify More DNA in Less Time

NucleoBond Xtra columns purify up to 2–5 times more DNA than other columns, in up to 60% less time. Plasmid DNA was isolated following each manufacturer's protocol, using the maximum culture volume and high plasmid content. Plasmid DNA yield was determined after DNA precipitation

Purify Endotoxin-Free DNA for Sensitive Cells

Endotoxins are components of the outer membrane of Gram-negative bacteria. They are toxic to cells grown in culture, and unfortunately they can be found in any non-sterile environment. NucleoBond Xtra EF kits use a patented procedure to reduce endotoxins to a very low level (0.05 EU/µg) that is suitable for the transfection of highly sensitive cells such as primary neurons and for lentivirus production.

Figure 2. A 9 kbp high-copy-number plasmid encoding a neuroglin-GFP fusion was purified using NucleoBond Xtra Midi EF and used to transfect primary rat hippocampal neurons. The neurons were analyzed by fluorescence microscopy 48 hr after transfection.

Simple Protocol to Purify Transfection-Grade DNA

The NucleoBond Xtra protocol is very simple: clear the bacterial lysate and load it onto the column, wash twice, and elute purified DNA in a high-salt buffer. Precipitate the purified DNA with isopropanol (Xtra kits) or for faster results, with the NucleoBond Xtra Finalizer (Xtra Plus kits). Endotoxin-free kits (NucleoBond Xtra EF and EF Plus) include one additional wash step.

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