Qiagen, 12945, QIAGEN Plasmid Plus Midi Kit (100)

100 QIAGEN Plasmid Plus Midi Columns, Extender Tubes, Reagents, Buffers, 100 QIAfilter Midi Cartridges

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 ").

Principle

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

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.

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

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

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