QIAGEN Plasmid Kits

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.

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Product Details
Specifications
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Cat No./ID: 10023

QIAGEN-tip 20 (25)

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25 columns

Cat No./ID: 10043

QIAGEN-tip 100 (25)

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25 columns

Cat No./ID: 10063

QIAGEN-tip 500 (25)

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25 columns

Cat No./ID: 10083

QIAGEN-tip 2500 (25)

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25 columns

Cat No./ID: 10091

QIAGEN-tip 10000 (5)

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5 columns

Cat No./ID: 12123

QIAGEN Plasmid Mini Kit (25)

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25 QIAGEN-tip 20, Reagents, Buffers

Cat No./ID: 12125

QIAGEN Plasmid Mini Kit (100).

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100 QIAGEN-tip 20, Reagents, Buffers

Cat No./ID: 12143

QIAGEN Plasmid Midi Kit (25)

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25 QIAGEN-tip 100, Reagents, Buffers

Cat No./ID: 12145

QIAGEN Plasmid Midi Kit (100)

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100 QIAGEN-tip 100, Reagents, Buffers

Cat No./ID: 12145X4

QIAGEN Plasmid Midi Kit (400)

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400 QIAGEN-tip 100, Reagents, Buffers

Cat No./ID: 12162

QIAGEN Plasmid Maxi Kit (10)

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10 QIAGEN-tip 500, Reagents, Buffers

Cat No./ID: 12163

QIAGEN Plasmid Maxi Kit (25)

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25 QIAGEN-tip 500, Reagents, Buffers

Cat No./ID: 12165

QIAGEN Plasmid Maxi Kit (100)

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100 QIAGEN-tip 500, Reagents, Buffers

Cat No./ID: 12165X4

QIAGEN Plasmid Maxi Kit (400)

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400 QIAGEN-tip 100, Reagents, Buffers

Cat No./ID: 12181

QIAGEN Plasmid Mega Kit (5)

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5 QIAGEN-tip 2500, Reagents, Buffers

Cat No./ID: 12183

QIAGEN Plasmid Mega Kit (25)

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25 QIAGEN-tip 2500, Reagents, Buffers

Cat No./ID: 12191

QIAGEN Plasmid Giga Kit (5)

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5 QIAGEN-tip 10000, Reagents, Buffers

Cat No./ID: 19046

Plasmid Buffer Set

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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.

Product Details

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.
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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.

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

Specifications

Features	Specifications
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

Product Resources

FAQs (50)

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	Is plasmid DNA purified with QIAGEN Plasmid Purification Kits suitable for in vitro transcription? FAQ ID -1	View
	When is chloramphenicol amplification of plasmids performed? FAQ ID -3	View
	What is the composition of buffer QBT? FAQ ID -411	View
	What is the composition of buffer QC? FAQ ID -412	View
	What is the composition of buffer QF? FAQ ID -413	View
	What is the composition of buffer STE? FAQ ID -415	View
	What is the composition of buffer TE? FAQ ID -416	View
	What is the composition of buffer P3? FAQ ID -418	View
	What can I do when the DNA pellet prepared with QIAGEN Plasmid Kits has been overdried? FAQ ID -572	View
	Where can I find a protocol for cleanup of already purified plasmid DNA? FAQ ID -1031	View
	I am seeing a precipitate after adding LyseBlue reagent to Buffer P1. What should I do about that? FAQ ID -1045	View
	What are the additional plasmid bands I see on my gel? FAQ ID -1059	View
	What is the composition of Buffer P2? FAQ ID -203	View
	How do I perform a DNA precipitation to concentrate my sample? FAQ ID -305	View
	Can Buffers N3 and P3 be used interchangeably? FAQ ID -310	View
	What is the white insoluble precipitate in my resuspended plasmid DNA pellet? FAQ ID -352	View
	Why do I get genomic DNA contamination in my plasmid prep? FAQ ID -353	View
	Can I eliminate RNase A from buffer P1 for my plasmid preparation to obtain RNase-free DNA for in-vitro transcription? FAQ ID -366	View
	Why is my plasmid DNA yield low? FAQ ID -768	View
	What is the advantage of running an analytical gel with fractions of my plasmid preparation? FAQ ID -769	View
	I left Buffer P1 at room temperature after addition of RNase A, what shall I do? FAQ ID -859	View
	What to do if cell clumps are present after Buffer P2 addition when using LyseBlue Reagent? FAQ ID -861	View
	Why would clumps occur following the addition of Buffer P2 when using LyseBlue Reagent in a plasmid preparation? FAQ ID -862	View
	With LyseBlue reagent for lysis control, can I now process more bacterial culture and overload the columns? FAQ ID -864	View
	Which QIAGEN plasmid preparation kits will contain LyseBlue Reagent? FAQ ID -865	View
	What is the expected level of endotoxins in plasmid DNA purified with QIAGEN Plasmid kits, QIAfilter Plasmid kits and HiSpeed Plasmid purification kits? FAQ ID -3082	View
	How do I know if my plasmid is a high- or low copy number type? FAQ ID -350	View
	What are the maximum culture volumes to use with the QIAGEN Plasmid Midi or Maxi Kit? FAQ ID -167	View
	What are the recommended culture and buffer volumes for a very low-copy plasmid? FAQ ID -168	View
	How should QIAGEN Plasmid Purification Kits be stored and for how long? FAQ ID -192	View
	Do you have a protocol for the isolation of very low-copy plasmids from Streptomyces spp.? FAQ ID -893	View
	Do you have a protocol for the isolation of single-stranded DNA from M13 phage using QIAGEN Plasmid Kits? FAQ ID -894	View
	What is the recipe for 2x YT? FAQ ID -213	View
	Do you have a protocol for the removal of endotoxins from already purified plasmid DNA? FAQ ID -500	View
	Do you have a protocol for the isolation of plasmid DNA from yeast? FAQ ID -853	View
	Do you have a protocol for the isolation of BAC DNA using the QIAGEN Plasmid Midi Kit? FAQ ID -881	View
	Do you have a protocol for the isolation of bacteriophage P1 derived constructs with any of your plasmid kits? FAQ ID -883	View
	Do you have a protocol for the isolation of plasmid DNA from Borrelia spp.? FAQ ID -884	View
	Do you have a protocol for the isolation of plasmid DNA from Citrobacter freundii? FAQ ID -885	View
	Do you have a protocol for the isolation of plasmid DNA from Lactobacillus spp.? FAQ ID -887	View
	Do you have a protocol for the isolation of plasmid DNA from Oligotropha carboxidovorans? FAQ ID -888	View
	Do you have a protocol for the isolation of plasmid DNA from Proteus spp.? FAQ ID -889	View
	Do you have a protocol for the isolation of plasmid DNA from Staphylococcus spp.? FAQ ID -890	View
	895 - Do you have a protocol for the isolation of endotoxin-free plasmid DNA using the QIAGEN Plasmid Midi Kit?	View
	What is the RNase A concentration and composition of Buffer P1? FAQ ID -198	View
	What is the recipe for LB? FAQ ID -212	View
	Do you have a protocol for the isolation of plasmid DNA from Bacillus subtilis? FAQ ID -896	View
	Do you have a protocol for the isolation of plasmid DNA from Corynebacterium glutamicum? FAQ ID -886	View
	Do you have protocols for the isolation of genomic DNA from tissue using QIAGEN-tips 2500 (Mega) and 10000 (Giga)? FAQ ID -907	View
	Do you have a protocol for the isolation of total RNA from plant tissue using QIAGEN-tips? FAQ ID -949	View

Kit Handbooks (1)

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	(EN) - QIAGEN Plasmid Purification Handbook — April 2012	Show details

Supplementary Protocols (8)

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	Re-Purification of Plasmid DNA Prepared by Methods other than QIAGEN Tips - (EN)	Show details
	Isolation of single-stranded DNA from M13 phage using QIAGEN® Plasmid Kits - (EN)	Show details
	Isolation of large-construct DNA using the QIAGEN® Plasmid Maxi Kit - (EN) This protocol is designed to provide up to 150 μg BAC/PAC/P1 DNA or up to 400 μg cosmid DNA.	Show details
	Isolation of genomic DNA from tissue using the QIAGEN-tip 10000 - (EN)	Show details
	Isolation of genomic DNA from tissue using the QIAGEN-tip 2500 - (EN) 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.	Show details
	Removal of endotoxins from purified plasmid DNA using the EndoFree® Plasmid Maxi Kit - (EN) Endotoxin-free DNA is essential for gene therapy research and will improve transfection into sensitive eukaryotic cells.	Show details
	Isolation of total RNA from plant tissue using the QIAGEN-tip - (EN) 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.	Show details
	Isolation of endotoxin-free plasmid DNA using the QIAGEN® Plasmid Midi Kit - (EN) This protocol is for purification of up to 100 µg endotoxin-free plasmid DNA using QIAGEN-tip 100.	Show details

Quick-Start Protocols (2)

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	QIAGEN Plasmid Mega and Giga Kits (EN)	Show details
	QIAGEN Plasmid Mini, Midi, and Maxi Kits (EN)	Show details

User-Developed Protocols (12)

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	Isolation of very low-copy plasmids from Streptomyces spp. using the QIAGEN® Plasmid Maxi Kit - (EN) 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.	Show details
	Isolation of BAC DNA using the QIAGEN® Plasmid Midi Kit - (EN) 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.	Show details
	Isolation of plasmid DNA from Bacillus subtilis using the QIAGEN® Plasmid Midi Kit - (EN) 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.	Show details
	Isolation of bacteriophage-P1-derived constructs using the QIAGEN® Plasmid Midi Kit - (EN) 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.	Show details
	Isolation of plasmid DNA from Borrelia spp. using the QIAGEN® Plasmid Midi Kit - (EN) 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.	Show details
	Isolation of plasmid DNA from Citrobacter freundii using the QIAGEN® Plasmid Midi Kit - (EN) 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.	Show details
	Isolation of plasmid DNA from Lactobacillus spp. using the QIAGEN® Plasmid Midi Kit - (EN) 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.	Show details
	Isolation of plasmid DNA from Oligotropha carboxidovorans using the QIAGEN® Plasmid Midi Kit - (EN) 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.	Show details
	Isolation of plasmid DNA from Proteus spp. using the QIAGEN® Plasmid Midi Kit - (EN) 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.	Show details
	Isolation of plasmid DNA from Staphylococcus spp. using the QIAGEN® Plasmid Midi Kit - (EN) 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.	Show details
	Isolation of plasmid DNA from yeast using the QIAGEN® Plasmid Midi Kit - (EN)	Show details
	Isolation of plasmid DNA from Corynebacterium glutamicum using the QIAGEN® Plasmid Mini Kit - (EN) 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.	Show details

Safety Data Sheets (19)

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	MSDS QIAGEN Plasmid Giga Kit (5)
	MSDS QIAGEN Plasmid Mega Kit (25)
	MSDS QIAGEN Plasmid Mega Kit (5)
	MSDS QIAGEN Plasmid Maxi Kit (100)
	MSDS QIAGEN Plasmid Maxi Kit (25)
	MSDS QIAGEN Plasmid Maxi Kit (10)
	MSDS QIAGEN Plasmid Midi Kit (100)
	MSDS QIAGEN Plasmid Midi Kit (25)
	MSDS QIAGEN Plasmid Mini Kit (100)
	MSDS QIAGEN Plasmid Mini Kit (25)
	MSDS QIAGEN-tip 10000 (5)
	MSDS QIAGEN-tip 2500 (25)
	MSDS QIAGEN-tip 500 (25)
	MSDS QIAGEN-tip 100 (25)
	MSDS QIAGEN-tip 20 (25)
	MSDS Plasmid Buffer Set
	MSDS QIAGEN Plasmid Maxi Kit (400)
	MSDS QIAGEN Plasmid Midi Kit (400)
	CofA

References

Product citations in scientific articles

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