产地:Qiagen货号:12943Applications The QIAGEN Plasmid Plus Midi Kit provides transfection-grade plasmid DNA, highly suited for all applications such as: Transfection into most cell lines (including sensitive cell lines such as Huh-7) High-quality automated sequencing Restriction digestion Cloning In vitro transcription/translation Principle and procedure The QIAGEN Plasmid Plus Midi Kit provides the most convenient method for large-scale plasmid preparation. The design and unique binding chemistry of the QIAGEN Plasmid Plus spin columns allow a simple bind-wash-elute procedure based on a novel chemistry. This procedure makes the QIAGEN Plasmid Plus Kit the fastest large-scale prep available - 20 minutes for 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. 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 in combination with the QIAvac holder) is used to draw the cleared lysate and subsequent wash buffers through the QIAGEN Plasmid Plus spin column. Following the washing and drying of the membrane, the highly concentrated DNA is eluted in low volumes by centrifugation (see figure "QIAGEN Plasmid Plus procedure"). 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 spin columns ensures that highly pure plasmid DNA is obtained every time. High-yield protocols and extra buffer volumes are provided with the QIAGEN Plasmid Plus Midi Kit, enabling yields of up to 250 μg of highly pure plasmid DNA, free of contaminants such as RNA, proteins, and genomic DNA (see figure "QIAGEN Plasmid Plus Kits (QPP) ensure high yields"). Transfection-grade plasmid DNA with very low endotoxin levels 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 kit reduces bacterial endotoxins to levels usually below 1 EU/μg* (see figures "Endotoxin levels when using QIAGEN Plasmid Plus Kits" and "Endotoxin levels (EU/μg DNA) using different plasmid DNA isolation methods"). This makes the plasmid DNA suitable for a multitude of applications, including transfection into sensitive cell lines (see figure "QIAGEN Plasmid Plus Kits show efficient transfection in sensitive cell lines"). 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.