QIAGEN Plasmid Plus Kits

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

S_1449_AppD_PLS1276

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QIAGEN Plasmid Plus Maxi Kit (25)

Cat. No. / ID:  12963

25 QIAGEN Plasmid Plus Maxi Columns, Extender Tubes, Reagents, Buffers, 25 QIAfilter Maxi Cartridges
KitAccessories
QIAGEN Plasmid Plus Kit
QIAvac Holder
VacValves
Column type
Midi
Maxi
Mega
Giga
Preparations
25
100
QIAGEN Plasmid Plus Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • 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

Product Details

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.

Want to try this solution for the first time? Request a quote for a trial kit.

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 1–3 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").

See figures

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").

See figures

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

Supporting data and figures

Resources

Technical Information (3)
Since QIAGEN launched the world’s first kit for purification of plasmid DNA, technologies have been developed to make plasmid DNA preparation even more convenient and reduce endotoxin levels in line with scientists’ needs in the laboratory. A lot of well-known companies have developed kits, and the choice of solutions available has never been greater. We compared popular advanced-technology kits to see their relative performance in speed, convenience of handling, yield and endotoxin levels.
Safety Data Sheets (1)
Brochures & Guides (1)
Certificates of Analysis (1)

FAQ

Can I use your QIAvac 96 for extraction of the DNA using QIAamp 96 DNA Blood Kit?

We do not recommend using a vacuum manifold for DNA extraction using the QIAamp 96 DNA Blood Kit because:

  1. Viscosity of the lysates varies from sample to sample. Therefore, the wells of the QIAamp 96 plate are at great risk of clogging.
  2. Use of vacuum manifold would result in incomplete removal of ethanol and salts after washing with Buffer AW2. This will affect the purity and yield of samples.
FAQ ID -9158
How can I purchase replacement bottles, caps and tubing for my QIAvac Connecting System?

Please contact Nalgene for ordering. Part numbers from Nalgene:

-          10 liter waste container (Nalgene Heavy Duty Vacuum Carboy [PP], cat. no. 2226-0020)

-          20 liter waste container (Nalgene Heavy Duty Vacuum Carboy [PP], cat. no. 2226-0050)

-          Quick filling/venting closure (Nalgene, cat. no. 2158-0021) [lid]

-          Vacuum tubing (e.g., Nalgene 180 clear plastic vacuum tubing, cat. no.8000-0065)

FAQ ID -9156
Which QIAGEN plasmid preparation kits will contain LyseBlue Reagent?

LyseBlue reagent is provided in the following QIAGEN plasmid kits:

FAQ ID -865
3313 - What is the endotoxin level for the QIAGEN Plasmid Plus kits?

Endotoxin level of QIAGEN Plasmid Plus kits is in the range of 1 EU/µg DNA.

 

What is the composition of Buffer S3?
The composition of Buffer S3 is confidential. This buffer is a proprietary component of QIAGEN Plasmid Plus Kits and is an plays and important role in the alkaline lysis procedure.  Buffer S3 can only be purchased in combination with QIAGEN Plasmid Plus Kits.
FAQ ID -2788
Can the VacConnectors for your QIAvac 24 Plus be reused?

VacConnectors prevent cross-contamination and are meant to be disposed after single use.

FAQ ID -9152
Can the waste bottles of the QIAvac Connecting System be autoclaved?

Yes, the waste bottles of the QIAvac Connecting System can be autoclaved. However, please be aware that the plastic may change in color.

FAQ ID -9153
3299-Can Buffer ER from the Endofree Plasmid kits be used instead of the Buffer ETR in QIAGEN Plasmid Plus kits?

No, the two buffers cannot be interchanged.  The QIAGEN Plasmid Plus chemistry is not compatible with Buffer ER from the Endofree Plasmid kits.

Can I use your QIAvac 24 Plus for extraction of DNA from tissues using the DNeasy Blood & Tissue Kit?

We do not recommend using a vacuum manifold for DNA extraction of tissues using the DNeasy Blood & Tissue Kit. Viscosity of the lysates varies from sample to sample; therefore, the DNeasy mini spin columns are at great risk of clogging.

FAQ ID -9154
Can the QIAvac 24 Plus manifold be autoclaved?

We do not recommend to autoclave the QIAvac 24 Plus manifold. Doing so can result in damage to the device.

FAQ ID -9151
Can I use your QIAvac 96 for extraction of DNA using the DNeasy 96 Blood & Tissue Kit?

We do not recommend using a vacuum manifold for DNA extraction with the DNeasy 96 Blood & Tissue Kit because:

  1. Viscosity of the lysates varies from sample to sample. Therefore, the wells of the DNeasy 96 plate are at great risk of clogging.
  2. Use of vacuum manifold would result in incomplete removal of ethanol and salts after washing with Buffer AW2. This will affect the purity and yield of samples.
FAQ ID -9157
Is the waste tray for the QIAvac Multiwell and the QIAvac 96 the same?

QIAvac Multiwell can hold plates that conform to SBS plate standards, whereas the QIAvac 96 does not. Therefore, the waste tray for QIAvac Multiwell is not the same as for our QIAvac 96.

However, the waste tray from QIAvac Multiwell will fit the QIAvac 96, but not vice versa.

FAQ ID -9159
The gasket on my QIAvac 96 seems to have cracks in it. Do you have a replacement part available, so I can replace the gasket?
We do not have this part available for replacement. You will need to replace your vacuum manifold. Please contact your Sales representative for a price quote.
FAQ ID -9161
The gasket on my QIAvac Multiwell doesn’t seem to seal. Do you have a replacement part available, so I can replace the gasket?
FAQ ID -9160
Can I use your QIAvac 24 Plus for extraction of DNA from tissues using the DNeasy Blood & Tissue Kit?

We do not recommend using a vacuum manifold for DNA extraction of tissues using the QIAamp Blood & Tissue Kit. Viscosity of the lysates varies from sample to sample; therefore, the DNeasy mini spin columns are at great risk of clogging.

FAQ ID -9155
What is the composition of Buffer BB?
The composition of Buffer BB is confidential. Buffer BB is a proprietary component of QIAGEN Plasmid Plus Kits and functions as a binding buffer, adjusting the correct binding conditions after alkaline lysis, when performing the QIAGEN Plasmid Plus procedure. Buffer BB can only be purchased in combination with QIAGEN Plasmid Plus Kits and is not available separately.
FAQ ID -2790
What is the composition of Buffer ETR?
The composition of Buffer ETR is confidential. Buffer ETR is a proprietary component of QIAGEN Plasmid Plus Kits and functions as an endotoxin removal buffer. Buffer ETR can only be purchased in combination with QIAGEN Plasmid Plus Kits.
FAQ ID -2789
Where can I buy the filter for QIAvac Connecting System?

The filter for QIAvac Connecting System is from Pall (part no. 4400). This is the Acro 50 filter, 0.2 µm PTFE, 1/8 inch MNPT connection. Please visit www.pall.com

FAQ ID -9150