16S rDNA amplicon sequencing of soil metagenome:bare soil with 12C-phenanthrene
Fragments of 430 bp covering the V3/V4 region of bacterial 16S rRNA genes were amplified by 28 cycles of PCR using Accuprime Super Mix (Invitrogen) on 1 l template DNA, purified using the UltraClean-htp 96 Well PCR Clean-Up kit (MOBIO) and quantified by Picogreen assay. An equimolar pool at 10 nM was purified using Nucleospin PCR Clean-Up kit (Macherey-Nagel) and sequenced on a single lane of Illumina Miseq PE250 at the Georgia Genomics Facility (Athens, GA, USA).