QIAGEN Plasmid Plus 96 Kits

For purification of up to 50 µg transfection-grade plasmid DNA in 96-well format

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QIAGEN Plasmid Plus 96 Miniprep Kit (4)

Cat. No. / ID:  16181

For 4 x 96 plasmid minipreps: TurboFilter 96 Plates, Plasmid Plus 96 Plates, Buffers, Reagents, S-Blocks, and Elution Microtubes; requires use of QIAvac 96 and Elution Microtube Adapter (available from QIAGEN Technical Service), or a centrifugation system suitable for 96-well blocks.
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ForFor instrument
QIAvac 96
BioRobot Universal
QIAGEN Plasmid Plus 96 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

  • Transfection-grade plasmid DNA with endotoxin levels <1 EU/µg DNA
  • DNA purity comparable to anion-exchange technology
  • High yields (up to 50 µg)
  • Convenient handling and simple procedure
  • Vacuum, centrifugation, and automated processing options

Product Details

QIAGEN Plasmid Plus 96 Kits provide a unique method for plasmid preparation in 96-well format. Samples can be conveniently processed using the QIAvac 96 and/or a centrifuge or automated on the BioRobot Universal System. Proprietary binding chemistry allows up to 50 µg of transfection-grade plasmid DNA per well to be obtained from up to 5 ml of an E. coli culture. Endotoxin levels are reduced to levels below 1 EU/µg DNA due to a dedicated endotoxin reduction buffer.
For optimal results it is recommended to use this product together with QIAvac 96.

Performance

Consistently high yields of ready-to-use plasmid DNA

QIAGEN Plasmid Plus technology delivers the same performance and quality as anion-exchange technology. High yields of up to 50 µg of transfection-grade plasmid DNA can be obtained (see figure " High yields of plasmid DNA").

Very low endotoxin levels

Kits include an endotoxin reduction buffer that effectively reduces bacterial endotoxins to levels typically lower than 1 EU/µg DNA, providing plasmid DNA quality which is suitable for transfection into sensitive cell lines (see figures " Very low endotoxin levels" and " Highly efficient transfection into a sensitive cell line").

See figures

Principle

QIAGEN Plasmid Plus 96 Kits are based on QIAGEN Plasmid Plus technology, a unique method for plasmid preparation. The procedure is based on a nonchaotropic binding chemistry that ensures selectivity and DNA quality that is comparable to anion exchange preps. Following lysate clearing, a simple bind-wash-elute procedure results in highly concentrated plasmid DNA, ready for direct use in subsequent applications. The unique kit chemistry and design of the Plasmid Plus 96 plates allow high binding capacities.

Procedure

This unique method for plasmid preparation in 96-well format provides highly pure, transfection-grade plasmid DNA using a simple bind-wash-elute procedure (see flowchart " QIAGEN PlasmidPlus 96 procedure").

The QIAGEN Plasmid Plus 96 Miniprep protocol starts with a modified alkaline lysis procedure. After neutralization, lysates are cleared using a TurboFilter 96 plate. A nonchaotropic binding buffer is added to the cleared lysate to optimize plasmid DNA binding to the membrane of the Plasmid Plus 96 plate. The unique binding buffer provides very specific binding conditions leading to DNA quality that is comparable to anion-exchange preps. A wash step with an endotoxin reduction buffer results in transfection-grade plasmid DNA suited for transfection into most cell lines (see figure " Highly efficient transfection into a sensitive cell lines"). The proprietary binding chemistry allows purification of up to 50 µg plasmid DNA per well from a 5 ml E. coli culture (see figure " High yields of plasmid DNA").

Convenient and flexible processing options

The procedure may be fully automated using the QIAGEN Plasmid Plus 96 BioRobot Kit on the BioRobot Universal System. Alternatively, the QIAvac 96 is used for lysate clearing using the TurboFilter 96 plate, and DNA binding and wash steps are performed using the Plasmid Plus 96 plate. Drying and elution steps may either be performed on the QIAvac 96 or using a compatible centrifuge. In addition, the complete procedure can be performed using a compatible centrifuge.

See figures

Applications

Plasmid DNA purified using QIAGEN Plasmid Plus 96 Kits is highly suitable for a wide variety of applications, including:

  • Transfection, including into sensitive cell lines
  • Preparation of short hairpin vectors (sh-vectors)
  • Enzymatic modifications
  • Cloning
  • Restriction digestion

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsTransfection into sensitive cell lines, preparation of short hairpin vectors (sh-vectors), enzymatic modifications, cloning, restriction digestion, sequencing, etc
Time per run or prep per run50 minutes
ProcessingQIAvac 96, centrifuge, automated
Culture volume/starting material1.25–5 ml culture volume
TechnologyQIAGEN Plasmid Plus technology
Yield15–50 µg
Samples per run (throughput)48–96 samples per run
Plasmid typeHigh-copy, low-copy, cosmid DNA

Resources

Kit Handbooks (1)
Quick-Start Protocols (1)
Certificates of Analysis (1)

FAQ

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.

 

For the the QIAprep 96 Plus Miniprep Kit or the QIAGEN Plasmid Plus 96 Miniprep Kit, can plates be partially used?

For the manual procedure, the plates can be partially used. Any unused wells should be sealed with tape. For the BioRobot Universal procedure, the entire 96 sample plate must be used. However, different sized plates may be available from our applications lab.  Please contact QIAGEN Technical Service for more details. 

https://www.qiagen.com/contact/contacttechnicalservice.aspx?WT.svl=m

FAQ ID -2319
Which kit, the QIAprep 96 Plus Miniprep Kit or the QIAGEN Plasmid Plus 96 Miniprep Kit, do you recommend for transfection?
For sensitive cell lines and downstream assays, we recommend QIAGEN Plasmid Plus 96 Miniprep Kit, because the endotoxin value is below 1 EU/µg DNA.  The QIAGEN Plasmid Plus 96 MiniPrep kit contains a special protocol for endotoxin removal.  Combination of other chemistries (such as Buffer ER) is not possible with this kit.  For robust cell lines, the QIAprep 96 Plus Miniprep Kit may be okay, as the endotoxin values are below 10 EU/µg DNA.
FAQ ID -2320
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.

What is the maximum plasmid size which I can use with the QIAprep 96 Plus Miniprep Kit or the QIAGEN Plasmid Plus 96 Miniprep Kit?
The QIAprep 96 Plus Miniprep Kit and the QIAGEN Plasmid Plus 96 Miniprep Kit can be used for the purification of plasmids up to 40 kb.
FAQ ID -2316
Can the QIAprep 96 Plus Miniprep Kit or the QIAGEN Plasmid Plus 96 Miniprep Kit be used for low-copy number plasmids?
The QIAprep 96 Plus Miniprep Kit and the QIAGEN Plasmid Plus 96 Miniprep Kit can be used for the purification of low-copy plasmids.
FAQ ID -2317
Are the QIAprep 96 Plus Miniprep Kit or the QIAGEN Plasmid Plus 96 Miniprep Kit suited for short hairpin vectors?
Yes, the QIAprep 96 Plus Miniprep Kit or the QIAGEN Plasmid Plus 96 Miniprep Kit are suited for the plasmid purification of short hairpin vectors?
FAQ ID -2318