For purification of up to 10 mg 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. Lysate clearing and isopropanol precipitation are achieved by centrifugation.
Buffers P1, P2, P3, QBT, QC, QF, RNase A; for 100 plasmid mini-, 25 midi-, or 10 maxipreps
QIAGEN Plasmid Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
Transfection efficiency versus plasmid purification method.
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.
QIAGEN Plasmid Kit procedures.
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.
Performance
The QIAGEN Plasmid Kits uses gravity-flow QIAGEN anion-exchange tips for efficient purification of plasmid DNA. Up to
10 mg (giga), 2.5 mg (mega), 500 µg (maxi), 100 µg (midi), and 20 µg (mini) high-copy plasmid DNA is purified from 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.
Specifications
Features
Plasmid
Giga Kit
Plasmid
Mega Kit
Plasmid
Maxi Kit
Plasmid
Midi Kit
Plasmid
Mini Kit
Applications
Transfection, cloning, sequencing, capillary sequencing, etc.
Transfection, cloning, sequencing, capillary sequencing, etc.
Transfection, cloning, sequencing, capillary sequencing, etc.
Transfection, cloning, sequencing, capillary sequencing, etc.
Transfection, cloning, sequencing, capillary sequencing, etc.
Culture volume/starting material
2.5–5 liters culture volume
500 ml – 2.5 liters culture volume
100–500 ml culture volume
25–100 ml culture volume
3–10 ml culture volume
Elution volume
Variable
Variable
Variable
Variable
Variable
Plasmid type
High-copy, low-copy, cosmid DNA
High-copy, low-copy, cosmid DNA
High-copy, low-copy, cosmid DNA
High-copy, low-copy, cosmid DNA
High-copy, low-copy, cosmid DNA
Processing
Manual (centrifugation)
Manual (centrifugation)
Manual (centrifugation)
Manual (centrifugation)
Manual (centrifugation)
Sample per run
1 sample per run
1 sample per run
1 sample per run
1 sample per run
1 sample per run
Technology
Anion-exchange technology
Anion-exchange technology
Anion-exchange technology
Anion-exchange technology
Anion-exchange technology
Time per run
320 min
220 min
160 min
150 min
80 min
Yield
<10 mg
<2.5 mg
<500 µg
up to 100 µg
<20 µg
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 suitable for use in applications, such as:
Transfection
Cloning
PCR
In vitro transcription
Features
Specifications
Applications
Transfection, cloning, sequencing, capillary sequencing etc.
