“…In order to allow specific degraders harboring the tfdA genes to proliferate, we treated the microcosms with 90 mol of 2,4-D, MCPA, or MCPP kg Ϫ1 of soil. Prior to and subsequent to Ն50% mineralization (measured using evolved 14 CO 2 ), a composite subsample consisting of 0.5 g of soil was removed, and DNA was extracted using a Power-Clean soil DNA kit (MoBio Laboratories, Carlsbad, CA) according to the manufacturer's instructions. Total DNA was quantified and the extraction efficiency was normalized by running 4-l aliquots of extracts on a standard 1.5% agarose gel stained with ethidium bromide, and qPCR was performed using the three different approaches (for further information on the experimental setup, nucleic acid preparation, and qPCR, see the supplemental Materials and Methods in the supplemental material).…”