Forced Labor Camp

The miniprep: One plasmid to billions

The miniprep: One plasmid to billions

Using a primitive microscope in 1600s, Robert Hooke observed and published the first images of cells. The discovery of cells was instrumental in understanding the role of microscopic things, from plant cells to bacteria. Bacteria govern the function of most ecosystems, but can also cause a number of diseases. For instance, the so-called 'Super Bug' MRSA (Methicillin-resistant Staphylococcus aureus) is a particular species of bacteria who's MO is feasting on the tiny skin cells of its victims. Humans and Nature have given them their super power of anti-biotic resistance, but that's another story. Escherichia coli (E. coli) is an unrelated bacterial species responsible for the bloody diarrhea associated with food poisoning, as well as a lot of cool biological procedures and eventual advancements in our understanding of how life works. Ecoli is hardy and replicates very fast (~20min) under the right conditions.

I perform a lot of minipreps using the E. coli strain DH5-a:

  • Wake up a vial of E. coli quaranteened at -80*C

  • Force feed them a plasmid I want more of

  • Let them multiply to billions of cells (22 hrs)

  • Crack them open

  • Extract and purify my now billions of plasmids for further applications

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( http://www.abnova.com ) - Miniprep plasmid DNA extraction kit is used when the starting E. coli culture volume is 1~5 ml of LB broth and the expected DNA yield is 20~30 μg. Other extraction and purification plasmid DNA Kits are available from varying manufacturers, named by size of bacterial culture, includes Gigaprep, Megaprep, and Midiprep.

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