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QIAGEN Plasmid Plus 96 Kits

For purification of up to 50 µg transfection-grade plasmid DNA in 96-well format
  • 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
QIAGEN Plasmid Plus 96 Kits provide a novel 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.
Cat No./ID: 16181
QIAGEN Plasmid Plus 96 Miniprep Kit (4)
$2,945.00
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For 4 x 96 plasmid minipreps: TurboFilter 96 Plates, Plasmid Plus 96 Plates, Buffers, Reagents, Flat-Bottom Blocks, S-Blocks, and Elution Microtubes; requires use of QIAvac 96 and Elution Microtube Adapter (available from QIAGEN Technical Services), or a centrifugation system suitable for 96-well blocks
Cat No./ID: 960241
QIAGEN Plasmid Plus 96 BioRobot Kit (4)
$2,983.00
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For 4 x 96 plasmid minipreps: TurboFilter 96 Plates and Plasmid Plus 96 Plates, Buffers, Reagents, Flat-Bottom Blocks, S-Blocks, and Elution Microtubes; for use with the BioRobot Universal System

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.


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Very low endotoxin levels.
pCMVβ plasmid DNA was purified from 1.3 ml and 5 ml E. coli DH10B cultures using the QIAGEN Plasmid Plus 96 Miniprep Kit, the QIAwell 96 Ultra Plasmid Kit, and kits from other suppliers. Endotoxin levels were determined using the Kinetic-QCL Kinetic Chromogenic Assay. For the EndoFree Plasmid Giga Kit, the endotoxin content was 0.0005 EU/μg DNA. EF: EndoFree Plasmid Giga Kit. QPP96: QIAGEN Plasmid Plus 96 Miniprep Kit. QW96: QIAwell 96 Ultra Plasmid Kit.
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High yields of plasmid DNA.
Using the QIAGEN Plasmid Plus 96 Miniprep Kit, pCMVβ plasmid DNA was purified from E. coli DH5a cultures grown in LB medium. DNA was quantified using absorbance measurement and verified by agarose gel analysis. High yields were achieved with the QIAGEN Plasmid Plus 96 Miniprep Kit.
2
Highly efficient transfection into a sensitive cell line.
pCMVβ DNA was prepared using the indicated preparation method. Huh-7 cells (4 x 104) were transfected using 200 ng plasmid DNA and 0.75 μl Attractene Transfection Reagent. β-gal activity and protein content were measured after 48 hours. Plasmid DNA prepared with QIAGEN Plasmid Plus 96 Miniprep and BioRobot Kits resulted in highly efficient transfection into sensitive cell lines.
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QIAGEN Plasmid Plus 96 procedure.
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").

Principle
QIAGEN Plasmid Plus 96 Kits are based on QIAGEN Plasmid Plus technology, a novel 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 novel 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 Plasmid Plus 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.

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
Features
Specifications
Applications Transfection into sensitive cell lines, preparation of short hairpin vectors (sh-vectors), enzymatic modifications, cloning, restriction digestion, sequencing, etc
Culture volume/starting material 1.25–5 ml culture volume
Plasmid type High-copy, low-copy, cosmid DNA
Processing QIAvac 96, centrifuge, automated
Samples per run; throughput 48–96 samples per run
Technology QIAGEN Plasmid Plus technology
Time per run or prep per run 50 minutes
Yield 15–50 µg

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Kit Handbooks (2)

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QIAGEN Plasmid Plus 96 Miniprep Kit
QIAGEN Plasmid Plus 96 BioRobot Kit
大腸菌からのトランスフェクション・グレードのプラスミドDNA 調製(96 ウェル)
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