About this item
The illustra™ plasmidPrep Mini Spin Kit uses a simple plasmid DNA purification protocol involving a modified alkaline lysis procedure and a novel silica-based membrane to achieve highly efficient plasmid DNA purification. Produces high quality plasmid DNA with excellent reproducibility for use in cloning, restriction enzyme digestion, PCR amplification, and DNA sequencing.
- Produces high-quality plasmid DNA with excellent reproducibility for use in cloning, restriction enzyme digestion, PCR amplification, and DNA sequencing
- Total preparation time of less than 10 min represents a 50% reduction in protocol time
- Increased ease of use provided by minimal changes in pipetting volume from one part of the protocol to the next
- Fewer centrifugation steps
- Color-coded tubes and columns simplify the protocol
- No organic solvents such as phenol chloroform
The extracted plasmid DNA resulting from performing a miniprep is called a "plasmid miniprep" and is often performed using a plasmid purification kit. Minipreps are used in the process of molecular cloning to analyze bacterial clones. When bacteria are lysed under alkaline conditions, both chromosomal DNA and protein are denatured, but the plasmid DNA remains stable. On addition of acetate-containing neutralization buffer the larger and less supercoiled genomic DNA and proteins precipitate, but the small bacterial DNA plasmids stay in the solution.
The typical plasmid DNA yield from a fresh 1.5 mL culture of E. coli containing a high copy number plasmid is approximately 9 μg. Instead of organic solvents, the kit contains chaotropic salts to denature protein components and promote selective binding of DNA to the novel silica membrane.
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