Molecular Biology Protocols

Diatomaceous Earth Purification of DNA-Mini, Midi, and Maxi Prep Methods

Note: In early 1994, there was much discussion on the bionet.molbio.methds-reagnts newsgroup regarding the efficacy and reproducibility of purification of DNA using Promega's Magic (now called Wizard) Preps. The technique below was submitted by Dr. Bruce Roe, Professor of Chemistry and Biochemistry at the University of Oklahoma (e-mail: broe@aardvark.ucs.uoknor.edu) to this newsgroup on February 17, 1994. Use of diatomaceous earth as a resin in DNA purification kits is evidently a patented procedure of Bio-Rad (Prep-a-gene)

Bruce Roe writes:

"I've been pretty quiet with all this discussion of Magic Mini preps. The recent re-posting of the "Merlin procedure" and at the request of several others, I am posting here our "cookbook" for using diatomaceous earth for maxi, midi, and mini-preps. The DNA prepared as described in this protocol (when followed by a capable scientist) results in DNA that works extremely well for sequencing (see comments in the protocol), restriction enzyme digestion and transformation. The DNA is stable when frozen and thawed a number of times and the Maxi prep. has been successfully used to isolate chimeric plasmids, cosmids and p1 constructs in the size range from 2.4kb to 100kb."

Note: Dr. Roe has now placed many of his lab's standard protocols on the Web. Click here to go to the Table of Contents of these methods.

Please reference:

K. Iyer, J. Crabtree, S. Clifton, S. Chissoe, E. Mardis and B. A. Roe, (1994) Isolation Of Cosmid, Plasmid and P1 DNAs via Diatomaceous Earth-based Maxi, Midi and Mini-preps, *and* the non-Diatomaceous Earth-based Mini-prep. manuscript submitted for publication.

Diatomaceous Earth Purification of DNA-Mini, Midi, and Maxi Prep Methods

IV. References.
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We have not used this protocol for isolating ss m13 templates as we previously published a robotic method for isolating 96 m13 templates in less than 3 hours and here are the references for the robotic m13 isolation procedure.