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Support >  Applications >  Nucleic_Acid_Purification >  Plasmid_DNA_Purification

Plasmid DNA Purification

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.

DNA purified with NucleoBond Xtra transfects efficiently

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

NucleoBond Xtra
Beats other Anion-Exchange Purification Columns
Midiprep Kit* Yield (µg) Time (min)
NucleoBond Xtra Plus* 520 28
Competitor Q HiSpeed* 335 47
NucleoBond Xtra 580 68
Competitor Q 260 185
Competitor I 380 131
NucleoBond Xtra
Beats Silica Membrane Purification, Too
Midiprep Kit Yield (µg) Time (min)
NucleoBond Xtra Plus* 520 28
Competitor S 300 37
Competitor P 90 41
* These kits use a desalting tool for 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.

Rat neurons transfected with endotoxin-free DNA.

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.

Simple plasmid DNA purification protocol

NucleoBond Xtra Specifications
  NucleoBond Xtra Midiprep NucleoBond Xtra Maxiprep
Applications Purify DNA for sequencing, cloning, and transfection
Technology Anion-exchange chromatography
Format Gravity flow columns
Starting Material < 200 ml bacterial lysate (high-copy-number plasmids) < 600 ml bacterial lysate (high-copy-number plasmids)
Lysate Clarification < 400 ml bacterial lysate (low-copy-number plasmids) < 1,200 ml bacterial lysate (low-copy-number plasmids)
Isopropanol Column filters
Precipitation Centrifugation (Midi kits)
NucleoBond Finalizer (Midi Plus kits)
Centrifugation (Maxi kits)
NucleoBond Finalizer (Maxi Plus kits)
Vector Size < 300 kb
Yield > 500 µg > 1,000 µg
Prep Time 68 min / 4 preps (Midi kits)
28 min / 4 preps (Midi Plus kits)
80 min / 4 preps (Maxi kits)
34 min / 4 preps (Maxi Plus kits)
Product Pages NucleoBond Xtra Midiprep Kits
Endotoxin-free NucleoBond Xtra Midiprep Kits
NucleoBond Xtra Maxiprep Kits
Endotoxin-Free NucleoBond Xtra Maxiprep Kits

Visit the Documents tab of the relevant product page to download the User Manual and other documentation.

 
 
Products
Cat. # Product Package Size Price # of Units Select
740410.10 NucleoBond® Xtra Midi 10 Preps $94.00
740410.100 NucleoBond® Xtra Midi 100 Preps $823.00
740412.10 NucleoBond® Xtra Midi Plus 10 Preps $114.00
740412.50 NucleoBond® Xtra Midi Plus 50 Preps $523.00
740414.10 NucleoBond® Xtra Maxi 10 Preps $217.00
740414.100 NucleoBond® Xtra Maxi 100 Preps $1,903.00
740414.50 NucleoBond® Xtra Maxi 50 Preps $1,021.00
740415 NucleoBond® Xtra Combi Rack Each $73.00
740416.10 NucleoBond® Xtra Maxi Plus 10 Preps $238.00
740416.50 NucleoBond® Xtra Maxi Plus 50 Preps $1,141.00
740417 NucleoBond® Xtra Buffer Set I Each $50.00
740410.50 NucleoBond® Xtra Midi 50 Preps $442.00
740420.10 NucleoBond® Xtra Midi EF 10 Preps $141.00
740420.50 NucleoBond® Xtra Midi EF 50 Preps $649.00
740422.10 NucleoBond® Xtra Midi Plus EF 10 Preps $177.00
740422.50 NucleoBond® Xtra Midi Plus EF 50 Preps $765.00
740424.10 NucleoBond® Xtra Maxi EF 10 Preps $296.00
740424.50 NucleoBond® Xtra Maxi EF 50 Preps $1,473.00
740426.10 NucleoBond® Xtra Maxi Plus EF 10 Preps $349.00
740426.50 NucleoBond® Xtra Maxi Plus EF 50 Preps $1,662.00
740436.10 NucleoBond® Xtra BAC 10 Preps $255.00
740436.25 NucleoBond® Xtra BAC 25 Preps $573.00
 

 

 



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