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.

Produto Product no. Num Cat. Preço de lista:
 
 
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Produto Num Cat. Preço de lista:
QIAGEN Plasmid Giga Kit (5)
5 QIAGEN-tip 10000, Reagents, Buffers
12191
Cotação
QIAGEN Plasmid Mega Kit (25)
25 QIAGEN-tip 2500, Reagents, Buffers
12183
Cotação
QIAGEN Plasmid Mega Kit (5)
5 QIAGEN-tip 2500, Reagents, Buffers
12181
Cotação
QIAGEN Plasmid Maxi Kit (100)
100 QIAGEN-tip 500, Reagents, Buffers
12165
Cotação
QIAGEN Plasmid Maxi Kit (25)
25 QIAGEN-tip 500, Reagents, Buffers
12163
Cotação
QIAGEN Plasmid Maxi Kit (10)
10 QIAGEN-tip 500, Reagents, Buffers
12162
Cotação
QIAGEN Plasmid Midi Kit (100)
100 QIAGEN-tip 100, Reagents, Buffers
12145
Cotação
QIAGEN Plasmid Midi Kit (25)
25 QIAGEN-tip 100, Reagents, Buffers
12143
Cotação
QIAGEN Plasmid Mini Kit (100)
100 QIAGEN-tip 20, Reagents, Buffers
12125
Cotação
QIAGEN Plasmid Mini Kit (25)
25 QIAGEN-tip 20, Reagents, Buffers
12123
Cotação
QIAGEN-tip 10000 (5)
5 columns
10091
Cotação
QIAGEN-tip 2500 (25)
25 columns
10083
Cotação
QIAGEN-tip 500 (25)
25 columns
10063
Cotação
QIAGEN-tip 100 (25)
25 columns
10043
Cotação
QIAGEN-tip 20 (25)
25 columns
10023
Cotação
Plasmid Buffer Set
Buffers P1, P2, P3, QBT, QC, QF, RNase A; for 100 plasmid mini-, 25 midi-, or 10 maxipreps
19046
Cotação
QIAGEN Plasmid Midi Kit (400)
400 QIAGEN-tip 100, Reagents, Buffers
12145X4
Cotação

QIAGEN Plasmid Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


 

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Transfection efficiency versus plasmid purification method.|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.
|
Desempenho
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.
Fundamento

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

Procedimento

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. 

Aplicações

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

  • Transfection
  • Cloning
  • PCR
  • In vitro transcription
Característica
Especificações
Applications Transfection, cloning, sequencing, capillary sequencing etc.
Culture volume/starting material 2.5–5 liters culture volume
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 320 min
Yield <10 mg

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Supplementary Protocols
8

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This protocol is designed to provide up to 150 μg BAC/PAC/P1 DNA or up to 400 μg cosmid DNA.
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This protocol is designed for the rapid, easy, and non-toxic preparation of up to 2 mg genomic DNA from not more than 2 g of tissue using QIAGEN-tip 2500. QIAGEN® Genomic-tips 20/G, 100/G, and 500/G can also be used with this protocol by reducing the amount of starting material according to the table on page 2. The purified genomic DNA ranges in size from 50-150 kb.
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Endotoxin-free DNA is essential for gene therapy research and will improve transfection into sensitive eukaryotic cells.
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This protocol is designed for isolation of up to 200 μg RNA from 150 mg plant tissue or up to 1 mg RNA from 600 mg plant tissue and is for use with QIAGEN-tip 100 or QIAGEN-tip 500, respectively.
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This protocol is for purification of up to 100 µg endotoxin-free plasmid DNA using QIAGEN-tip 100.
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User-Developed Protocols
12
The procedure has been used successfully for isolation of plasmids SCP2 and SCP2* as well as the plasmids listed in Table 1 (see next page) from S. coelciolor and S. lividans strains.
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This procedure has been used successfully for isolation of 150-250 kb BAC DNA from a mouse-BAC library cloned in pBeloBAC11 from Escherichia coli strain HB101/r. The yield of BAC DNA from 100 ml culture was typically 20-40 μg.

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The procedure has been used successfully for isolation of high- and low-copy-number plasmids from various Bacillus subtilis strains. Yield of plasmid DNA was typically 10-20 µg plasmid DNA from 100 ml culture.
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The procedure has been used successfully for isolation of 110 kb P1 DNA (pAdsacBII with an 80 kb insert) from Escherichia coli strain NS3529. Yield of P1 DNA was typically 10-50 µg from 500 ml culture.
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The procedure has been used successfully for isolation of linear plasmids from Borrelia burgdorferi sensu lato species, which include Borrelia burgdorferi sensu stricto, Borrelia afzelli, and Borrelia garinii.
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The procedure has been used successfully for isolation of high-copy-number plasmids from Citrobacter freundii. Yield of plasmid DNA was typically 3-8 µg DNA per ml culture.
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The procedure has been used successfully for isolation of cryptic plasmids (pLC2-based) from mesophilic Lactobacillus strains such as L. sake and L. curvatus. Yield of plasmid DNA was typically 10-20 µg plasmid DNA from 100 ml culture.
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The procedure has been used successfully for isolation of the large (128 kb), very-low-copynumber (1-2 copies per cell) plasmid pHCG3 and its derivatives from Oligotropha carboxidovorans. Yield of plasmid DNA was typically 3-6 µg plasmid DNA from 200 ml culture.
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The procedure has been used successfully for isolation of high-copy-number plasmids from Proteus vulgaris and Proteus mirabilis. Yield of plasmid DNA was typically 3-8 µg DNA per ml culture.
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The procedure has been used successfully for isolation of a variety of medium-copy-number shuttle vectors from S. xylosus, S. carnosus, S. epidermidis, and S. aureus. Yield of plasmid DNA was typically 2-10 µg from 50 ml culture.
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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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Referências
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