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.
Holder for stabilizing QIAGEN Plasmid Plus Mega and Giga columns with Tube Extenders on the QIAvac 24 Plus
QIAGEN Plasmid Plus Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
Efficient transfection in sensitive cell lines.
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.
Endotoxin levels using QIAGEN Plasmid Plus Kits.
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.
Endotoxin levels using different isolation methods.
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.
High yields are ensured with QIAGEN Plasmid Plus Kits (QPP).
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.
Analytical chromatogram.
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.
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
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
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
Features
Specifications
Applications
Transfection of most cell lines, cloning, enzymatic restriction, in-vitro translation/transcription, sequencing
Culture volume/starting material
50–200 ml culture volume
Final DNA concentration
≥ 1 µg/µl
Plasmid type
Plasmids, cosmids
Processing
manual (centrifugation and vacuum), parallel processing
For 50 preps: QIAamp Mini Columns, Tube Extenders (20 ml), QIAGEN Proteinase K, Carrier RNA, Buffers, VacConnectors, and Collection Tubes (1.5 ml and 2 ml)
2.5 ml (100 mg/ml; 7000 units/ml, solution)
Unit definition: That amount of enzyme causing the hydrolysis of RNA at a rate such that k (velocity constant) equals unity (Kunitz units) at 25°C and pH 5.0.
