Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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Preparation of Plasmid DNA by Alkaline Lysis with SDS: Maxipreps

(Protocol summary only for purposes of this preview site)

In recent years, with the advent of PCR and the development of highly efficient methods of cloning and DNA sequencing, the need to prepare large quantities of plasmid vectors and recombinants has greatly diminished. In consequence, this protocol, at one time in common use, has been largely replaced by faster and easier column-based purification methods (see the discussion of Commercial Kits for Purification of DNA in the chapter introduction). In this protocol, a scaled-up version of the alkaline lysis method (Birnboim and Doly 1979), plasmid DNA is purified from the cleared bacterial lysate by centrifugation to equilibrium in CsCl gradients containing ethidium bromide. These gradients have an aesthetic quality that justifies the continued inclusion in this manual of a protocol that in many other respects is an antique.


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(Limited time special offer.) Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook
Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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