For the fastest, most convenient purification of up to 10 mg transfection-grade plasmid DNA

  • Rapid purification of up to 24 samples
  • Fast vacuum-based parallel processing
  • Low elution volumes for highly concentrated DNA
  • Very low endotoxin levels (<1 EU/μg) due to dedicated wash buffer

The QIAGEN Plasmid Plus Kits enable ultrafast, large-scale purification of up to
10 mg of highly pure plasmid DNA. The use of a vacuum manifold allows purification of up to 24 samples in parallel, reducing hands-on time. Low elution volumes yield highly concentrated plasmid DNA for direct use without ethanol precipitation. The QIAGEN Plasmid Plus Kits also feature a dedicated wash buffer for endotoxin reduction. The plasmid DNA obtained is highly suitable for a multitude of applications, including transfection into sensitive cell lines.
For optimal results it is recommended to use this product together with QIAvac 24 Plus.

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Product Cat. no. List price:
QIAGEN Plasmid Plus Giga Kit (5)
5 QIAGEN Plasmid Plus Mega Columns, Extender Tubes, Reagents, Buffers, 5 QIAfilter Mega-Giga Cartridges
12991
464,00 €
QIAGEN Plasmid Plus Mega Kit (5)
5 QIAGEN Plasmid Plus Mega Columns, Extender Tubes, Reagents, Buffers, 5 QIAfilter Mega-Giga Cartridges
12981
221,00 €
QIAGEN Plasmid Plus Maxi Kit (100)
100 QIAGEN Plasmid Plus Maxi Columns, Extender Tubes, Reagents, Buffers, 100 QIAfilter Maxi Cartridges
12965
1 982,00 €
QIAGEN Plasmid Plus Maxi Kit (25)
25 QIAGEN Plasmid Plus Maxi Columns, Extender Tubes, Reagents, Buffers, 25 QIAfilter Maxi Cartridges
12963
526,00 €
QIAGEN Plasmid Plus Midi Kit (100)
100 QIAGEN Plasmid Plus Midi Columns, Extender Tubes, Reagents, Buffers, 100 QIAfilter Midi Cartridges
12945
883,00 €
QIAGEN Plasmid Plus Midi Kit (25)
25 QIAGEN Plasmid Plus Midi Columns, Extender Tubes, Reagents, Buffers, 25 QIAfilter Midi Cartridges
12943
245,00 €
QIAvac Holder
Holder for stabilizing QIAGEN Plasmid Plus Mega and Giga columns with Tube Extenders on the QIAvac 24 Plus
19418
113,00 €
VacValves (24)
24 valves for use with the QIAvac 24 and QIAvac 24 Plus
19408
104,00 €
QIAGEN Plasmid Midi Kit (400)
400 QIAGEN-tip 100, Reagents, Buffers
12145X4
2 511,00 €

QIAGEN Plasmid Plus 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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Efficient transfection in sensitive cell lines.|Endotoxin levels using QIAGEN Plasmid Plus Kits.|Endotoxin levels using different isolation methods.|High yields are ensured with QIAGEN Plasmid Plus Kits (QPP).|QIAGEN Plasmid Plus procedure.|Analytical chromatogram.|
Plasmid pCMVβ DNA was prepared using the indicated preparation method with standard and high-yield (HY) protocol for QIAGEN Plasmid Plus Kits. Huh-7 cells were transfected using 200 ng plasmid DNA and 0.5 μl Attractene Transfection Reagent or 300 ng plasmid DNA and 0.75 μl Attractene Transfection Reagent.|Purification per pellet-wet weight (g/l) for midi prep using Buffer ETR is shown. QIAGEN Plasmid Plus Technology generally results in low endotoxin levels. Buffer ETR technology further decreases these levels.|QIAGEN Plasmid Plus Kits result in lower EU content levels than kits based on silica and anion-exchange technology from other suppliers. For EndoFree Plasmid Kits, the EU content is below detection range.|Comparison of plasmid DNA yields using different methods — QIAGEN Plasmid Plus high-yield protocol (QPP), QIAGEN Plasmid Plus standard protocol (QPP), QIAGEN Plasmid Kit (QP), Supplier I, and Supplier S — is shown. Microgram DNA/ml biomass is indicated.|QIAGEN Plasmid Plus Kits provide a rapid and convenient method for large-scale plasmid preparation.|Plasmid DNA (100 μg) purified using the QIAGEN Plasmid Plus Mega Kit was analyzed using HPLC on an analytical anion-exchange column to detect non-plasmid nucleic acid contamination (i.e., RNA or genomic DNA). No contamination is present in the plasmid prep.
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Performance
Consistent quality and performance

QIAGEN Plasmid Plus technology delivers the same performance and quality as anion-exchange technology. The unique binding chemistry of the QIAGEN Plasmid Plus columns ensures that pure plasmid DNA is obtained every time. High-yield protocols and extra buffer volumes are provided with the QIAGEN Plasmid Plus Kits. Yields of up to 10 mg (giga),
2 mg (mega), 1 mg (maxi), and 250 μg (midi) of highly pure plasmid DNA, free of contaminants such as RNA, proteins, and genomic DNA are achieved (see figures "High yields are ensured with QIAGEN Plasmid Plus Kits (QPP)"  and "Analytical chromatogram").

Transfection-grade plasmid DNA with very low endotoxin levels

Endotoxins, also known as lipopolysaccharides or LPS, are cell-membrane components of Gram-negative bacteria such as E. coli and are released during the lysis step of plasmid purification. Endotoxins can induce nonspecific activation of immune responses, and thus represent a noncontrollable variable in transfection experiment setup, influencing the outcome and reproducibility of results and making them difficult to compare and interpret.  

The endotoxin wash buffer provided in the kits reduces bacterial endotoxins to levels usually below 1 EU/μg DNA. (Endotoxin levels can vary depending on the host strain, culture media, and cell density. If the cell density is very high, endotoxin levels may be slightly higher, ranging from 13 EU/μg DNA.) This makes the plasmid DNA suitable for a multitude of applications, including transfection into sensitive cell lines (see figures "Endotoxin levels using QIAGEN Plasmid Plus Kits", "Endotoxin levels using different isolation methods", and "Efficient transfection in sensitive cell lines"). 

Principle

The QIAGEN Plasmid Plus Kits provide the most convenient method for large-scale plasmid preparation. The design and unique binding chemistry of the QIAGEN Plasmid Plus columns allow a simple bind-wash-elute procedure based on a novel chemistry. This procedure makes these kits the fastest large-scale prep available 20 minutes for maxi and midi preps. The resulting plasmid DNA is highly concentrated and ready for immediate use in subsequent applications without further ethanol precipitation. Thus, the plasmid DNA is highly suitable for applications that only allow small reaction volumes.

Maximum plasmid yields using LyseBlue

LyseBlue, an optional color indicator that provides visual identification of optimum buffer mixing, is also included in the kit. This prevents common handling errors that lead to inefficient cell lysis and incomplete precipitation of SDS, genomic DNA, and cell debris. Through a simple color change, this unique buffer additive ensures that optimum cell lysis and neutralization is achieved, maximizing yields.

Specifications

Features
Plasmid Plus
Giga Kit
Plasmid Plus 
Mega Kit
Plasmid Plus 
Maxi Kit
Plasmid Plus 
Midi Kit
Applications Transfection of most cell lines, cloning, enzymatic restriction,
in-vitro translation/transcription, sequencing
Transfection of most cell lines, cloning, enzymatic restriction,
in-vitro translation/transcription, sequencing
Transfection of most cell lines, cloning, enzymatic restriction,
in-vitro translation/transcription, sequencing
Transfection of most cell lines, cloning, enzymatic restriction,
in-vitro translation/transcription, sequencing
Culture volume/starting material 2500 ml culture volume 500 ml culture volume 50–200 ml culture volume 25–50 ml culture volume
Final DNA concentration ≥1 µg/µl ≥1 µg/µl ≥1 µg/µl ≥1 µg/µl
Plasmid type Plasmids, cosmids Plasmids, cosmids Plasmids, cosmids Plasmids, cosmids
Processing Manual (centrifugation and vacuum), parallel processing Manual (centrifugation and vacuum), parallel processing Manual (centrifugation and vacuum), parallel processing Manual (centrifugation and vacuum), parallel processing
Samples per run; throughput 1–12 samples per run 1–12 samples per run 1–24 samples per run 1–24 samples per run
Technology Advanced silica technology (non-chaotrophic), endotoxin reduction Advanced silica technology (non-chaotrophic), endotoxin reduction Advanced silica technology (non-chaotrophic), endotoxin reduction Advanced silica technology (non-chaotrophic), endotoxin reduction
Time per run 50 min 40 min  20 min 20 min
Yield up to 10 mg up to 2.5 mg up to 1 mg up to 250 µg

Procedure
QIAfilter cartridges provided in QIAGEN Plasmid Plus Kits enable rapid lysate clearing. Loading of the cleared lysate onto the column can be performed in one step, saving time. A vacuum manifold (such as the QIAvac 24 Plus) is used to draw the cleared lysate and subsequent wash buffers through the QIAGEN Plasmid Plus column. Following the washing and drying of the membrane, the highly concentrated DNA is eluted in low volumes by centrifugation (see flowchart "QIAGEN Plasmid Plus procedure").
Applications

The QIAGEN Plasmid Plus Kits provide transfection-grade plasmid DNA highly suited for all applications, such as:

Transfection into most cell lines (e.g., sensitive cell lines such as Huh-7)
Restriction digestion
Cloning
In vitro transcription and translation
Sequencing

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