For purification of up to 20 µg transfection-grade plasmid or cosmid DNA

  • Purity equivalent to 2 x CsCl gradient centrifugation
  • High yields of plasmid DNA
  • Cost-effective preparations
  • LyseBlue for optimum lysis and maximum DNA yield

QIAGEN Plasmid Kits provide gravity-flow, anion-exchange tips for purification of transfection-grade plasmid DNA. The QIAGEN Plasmid Mini Kit delivers DNA yields of up to 20 μg. Lysate clearing and isopropanol precipitation are achieved by centrifugation.

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Quantity Product Cat. no. Price Sum
 
QIAGEN Plasmid Mini Kit (25)
25 QIAGEN-tip 20, Reagents, Buffers
12123
$200.00
$0.00
 
QIAGEN Plasmid Mini Kit (100)
100 QIAGEN-tip 20, Reagents, Buffers
12125
$638.00
$0.00
The QIAGEN Plasmid Mini Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Transfection efficiency versus plasmid purification method.|Comparison of QIAGEN Plasmid Kit procedures.|
Different pRSVcat DNA preparations using the methods indicated were introduced into the indicated cell lines by liposome-mediated transfection, and the efficiencies determined by measuring CAT expression levels after 40 h. Each bar represents the mean of 4 independent transfections (2 transfections with each of 2 independent plasmid preparations). The highest transfection efficiency was achieved with QIAGEN Plasmid Kits.
|Neutralized bacterial lysates are cleared by centrifugation. The cleared lysate is then loaded onto the anion-exchange tip where plasmid DNA selectively binds under appropriate low-salt and pH conditions. RNA, proteins, metabolites, and other low-molecular-weight impurities are removed by a medium-salt wash, and ultrapure plasmid DNA is eluted in high-salt buffer. The DNA is concentrated and desalted by isopropanol precipitation and collected by centrifugation.
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Performance
Classic QIAGEN Plasmid Mini Kits use gravity-flow QIAGEN-tip 20 anion-exchange tips for efficient purification of plasmid DNA. Up to 20 µg high-copy plasmid DNA is purified from 3–10 ml culture. (Culture volumes depend on plasmid copy number, size of insert, host strain, and culture medium.) Plasmid DNA purified with QIAGEN Plasmid Kits is highly suitable for use in applications such as transfection (see figure "Transfection efficiency versus plasmid purification method"), cloning, and in vitro transcription.
Principle

The unique anion-exchange resin in QIAGEN-tips is developed exclusively for the purification of nucleic acids. Its exceptional separation properties result in DNA purity equivalent or superior to that obtained by two successive rounds of CsCl gradient centrifugation. Prepacked QIAGEN-tips operate by gravity flow and never run dry, minimizing the hands-on time required for plasmid preparation. The entire QIAGEN plasmid purification system avoids the use of toxic substances such as phenol, chloroform, ethidium bromide, and CsCl, minimizing hazard both to the user and the environment.

Procedure

With QIAGEN Plasmid Kits, bacterial lysates are cleared by centrifugation. The cleared lysate is then loaded onto the anion-exchange tip where plasmid DNA selectively binds under appropriate low-salt and pH conditions. RNA, proteins, metabolites, and other low-molecular-weight impurities are removed by a medium-salt wash, and pure plasmid DNA is eluted in high-salt buffer (see flowchart "QIAGEN Plasmid Kit procedures"). The DNA is concentrated and desalted by isopropanol precipitation and collected by centrifugation.

Applications

Plasmid DNA purified with QIAGEN Plasmid Kits is highly suited for use in applications, such as:

Transfection
Cloning
PCR
In vitro transcription
Feature
Specifications
Applications Transfection, cloning, sequencing, capillary sequencing etc.
Culture volume/starting material 3–10 ml culture volume
Elution volume Variable
Plasmid type High-copy, low-copy, cosmid DNA
Processing Manual (centrifugation)
Samples per run; throughput 1 sample per run
Technology Anion-exchange technology
Time per run or prep per run 80 min
Yield <20 µg

FAQs
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User-Developed Protocols
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The procedure has been used successfully for isolation of different medium-copy-number plasmids carrying pHM1519 or pBL1 origins of replication from Corynebacterium glutamicum ATCC 13032. Yield of plasmid DNA was typically 0.4-1.5 µg per ml LB culture, although yield was dependent on the vector, the insert, and the size of the plasmid.
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