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. The method previously used in our laboratory 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 × 1010 to 2–4 × 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.
Promega Notes 68, 30.
Shermalyn R. Greene and Lola V. Stamm