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."
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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169-182.
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Grossman, L. and Moldave, K., Eds. )29: 363-418.
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alkaline extraction method for the isolation of plasmid DNA. Methods in
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Clewell, D.B., and D.R. Helinski. 1969. Supercoiled circular DNA-protein complex
in E.coli: Purification and induced conversion to an open circular DNA form.
Proc. Natl.Acad.Sci.U.S.A. 62:1159-1166.
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L. Pinck. 1989. A rapid and efficient 'miniprep' for the isolation of plasmid
DNA. Nucl. Acids. Res. 17: 3604.
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preparation yielding DNA suitable for double stranded sequencing and in Vitro
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of plasmid DNAs by Hydroxyapatite chromatography. Eur.J.Biochem. 91: 303-310.
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135:345-348.
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Purification of RNA free plasmid DNA using alkaline extraction followed by
Ultrogel A2 column chromatography. Anal.Biochem. 148: 121-126.
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R. Chipperfield and H.C. Birnboim. 1982. A procedure for the large scale
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1979. Preparative and analytical purification of DNA from agarose.
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method for purification of nucleic acids. Jour. of Clin. Micro. 28: 495-503.
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purified plasmid DNA from Bacillus subtilis. Applied and Environmental
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inexpensive and simple method for DNA purifications on silica particles.
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Little. 1990. Prep-A-GeneTM: A superior matrix for the purification of DNA and
DNA fragments. Biotechniques 9:92-99.
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chromatography of nucleic acid components (Isolation of the minor nucleosides).
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Noyer-Weidner, S.S. Smith, M.Z. Michael and M.W. Graham. 1989. Quantitative
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Walker and R.R. McInnes. 1993. E.coli host strains significantly affect the
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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.
- E.R. Mardis
and B.A. Roe. Automated Methods for Single-Stranded DNA Isolation and
Dideoxynucleotide DNA Sequencing Reactions on a Robotic Workstation.
BioTechniques 7, 840-850 (1989).
- S. L. Chissoe, Y.F. Wang, S.W. Clifton, N. Ma, H.J. Sun, J. S. Lobsinger, S.
M. Kenton, J. D. White and B. A. Roe. Strategies for Rapid and Accurate DNA
Sequencing. Methods: A Companion to Methods In Enzymology. 3, 55-65 (1991).