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Efficient Recovery of Ultrapure Plasmid DNA

s those produced using endotoxin-free kits (Fig. 3).

Quality controls

To ensure that they always meet the requirements for reproducible quality and quantity, Eppendorf plasmid kits are subject to stringent quality-control procedures. Control plasmids (high-/low-copy) are isolated in accordance with protocol guidelines, and their quality and quantity is then determined using a variety of methods, including spectrophotometric assays,3 agarose gel electrophoresis, restriction digestion, and automatic fluorescence sequencing. Transfection of culture cells via calcium phosphate provides an additional, and important, proof of quality.

Fig. 2: The concentration of plasmid DNA obtained with the plasmid purification kit from Eppendorf and that obtained with a commonly available kit (mini- and maxi-prep scale). In each case, the concentration was determined spectrophotometrically. Midi-Prep 1 (25 ml preparation): Vector pEGFP-N1; length: 4.7 kb; bacteria strain: HB-101; Midi-Prep 2 (75 ml preparation): Vector pCH110; length: 7.128 kb; bacteria strain: JM-109; Maxi-Prep 1 (100 ml preparation): Vector pCH110; bacteria strain: JM-109; Maxi-Prep 2 (100 ml preparation): Vector pGEMgapdh; length: 2.7 kb; bacteria strain: HB-101. Each experiment was carried out twice. Fig. 3: Comparison between the transfection efficiency of the Maxi plasmid purification kit from Eppendorf (ENH) and that of a commonly available kit (A) and a kit that can be used to extract endotoxin-free DNA (B). Positive transfections were detected via FACS
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