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Isolation
of Spirochete DNA Using the Wizard® Genomic DNA Purification Kit
By Shermalyn R. Greene, Ph.D., and
Lola V. Stamm, Ph.D.
Department of Epidemiology, University of North Carolina, Chapel Hill
Purified genomic DNA can be isolated
rapidly from the spirochetes Treponema pallidum (syphilis agent) and T.
denticola (periodontal disease agent) using the Wizard® Genomic DNA
Purification System (1-3). The method previously used in our laboratory (4) involved
organic (phenol/chloroform/isoamyl alcohol) extraction and required approximately three
days to complete. The method described here requires 1-2 hours and consistently achieves
higher DNA yields. Higher genomic DNA yields are particularly valuable for our work with
T.
pallidum, because this species must be cultured in vivo, producing only limited
amounts of starting material. Using the Wizard® Genomic Kit for Treponema
species, we have dramatically reduced input cell numbers from 1-2 x 1010 to 2-4
x 109. Yields are 0.8-1µg/µl in 100µl. Additional advantages to this method
are its greater simplicity and the elimination of hazardous chemicals and concerns of
accompanying waste disposal.
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