Rapid Plasmid Purification Kits, Rapid Isolation of Plasmid DNA, Marligen PowerPrep Express Plasmid Purification

Marligen’s PowerPrep™ Express Plasmid Purification Kits are designed for the rapid isolation of plasmid DNA. The procedure involves modified alkaline lysis and binding to high capacity silica membranes followed by elution with TE or water. The purified DNA is ready to use for most standard molecular biology applications including restriction mapping, cloning, PCR amplification, labeling and DNA sequencing. Columns are available in miniprep, midiprep and maxiprep sizes for use with either vacuum (mini only) or centrifugation-based methods.

Technology

A modified alkaline lysis is used to create a cleared lysate containing plasmid DNA with minimal amounts of genomic DNA. The lysate is applied to a spin cartridge containing high capacity silica membranes to which the plasmid DNA binds. Using optimized protocols and buffers, the contaminants are washed away and the highly purified plasmid DNA is eluted in TE buffer or water, ready to use.

(Note: for purifying low copy number plasmids or for preparing large insert clones, please see Marligen’s PowerPrep™ HP Plasmid Purification Kits).

Performance Characteristics

DNA purified with Marligen’s PowerPrep™ Express Plasmid Purification Kits is suitable for most applications in molecular biology. Figure 1 shows the excellent reproducibility obtained with Marligen’s PowerPrep™ Express Miniprep Kit. Figure 2 demonstrates complete cutting of plasmid DNA with a variety restriction enzymes. Figure 3 shows that plasmid DNA purified with Marligen’s PowerPrep™ Express Maxiprep Kit yields DNA that can be re-ligated to completion following digestion with EcoR1. Purity, integrity, performance in restriction endonuclease digestion and ligation, and column capacity is routinely tested on each lot.

twelve replicate purifications of the plasmid pT7T3

Fig. 1 Twelve replicate purifications of the plasmid pT7T3 containing a 1.2 kb insert were performed with the miniprep size of the Marligen PowerPrep™ Express Plasmid Purification System. The results demonstrate a very high level of reproducibility.

restriction digestion of plasmid DNA purified with the marligen rapid plasmid purification system

Fig. 2 Restriction digestion of plasmid DNA purified with the Marligen PowerPrep™ Express Plasmid Purification System. Lane 1 shows the undigested plasmid DNA and lane 2 shows plasmid DNA that has been linearized with BamH1. The remaining digestions were performed with Bgll (lane 3), Ddei (lane 4), Dra1 (lane 5), EcoR1 (lane 6), HaeIII (lane 7), and HindIII (lane 8).

plasmid dna prepared with the marligen rapid plasmid purification system

Fig. 3 Plasmid DNA prepared with the Marligen PowerPrep™ Express Plasmid Purification System was digested with EcoR1 and ligated with a 1.7 kb DNA fragment. Lane 2 shows the DNA before ligation. Ligations were performed with 0.03 U (lane 3), 0.06 U (lane 4), 0.08 U (lane 5), 0.10 U (lane 6) and 0.12 U (lane 7) of T4 DNA Ligase per ug of DNA.

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