Supercoiled pBR322 DNA

Supercoiled plasmid pBR322 DNA is produced by the large-scale alkaline-lysis method (Sambrook et al., 1989)¹ from a high-copy derivative of pBR322 (Boros et al., 1984)².

Negatively-supercoiled pBR322 DNA is further treated so that the DNA is more supercoiled and has a narrow range of linking numbers making it an ideal substrate for relaxation reactions.

Positively-supercoiled plasmid pBR322 DNA is produced by treating relaxed pBR322 with reverse gyrase before final purification.

It is supplied at a concentration of 1mg/ml in TE (10mM Tris-HCl (pH 7.5), 1mM EDTA). Store at 4^oC

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References

1. Sambrook, J., Fritsch, E.F. & Maniatis, T (1989). Molecular cloning: a laboratory manual,2nd ed., Cold Spring Harbor Press. Cold Spring Harbor, NY.
2. Boros, I., Pósfai, G. & Venetianer, P. (1984). High-copy number derivatives of the plasmid cloning vector pBR322. Gene 30, 257-260