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Genomic DNA was extracted using the UltraClean? Soil DNA Isolation kit (MoBio Laboratories Inc., Carlsbad, CA). Bacterial 16S rRNA genes were amplified from DNA extracts using the universal bacterial primers 27f (5�� AGA GTT TGA TCM TGG CTC AG 3��) and 1492f (5�� TAC GGY TAC CTT GTT ACG ACT T 3��) (Lane et?al., 1991). For T-RFLP analysis, 27f primers were 5��-end labeled with WELLRED D4 (Sigma-Proligo; Sigma-Aldrich Canada Ltd., Oakville, ON, Canada). Amplification was performed by an initial denaturation step at 94?��C for 4?min, 30 cycles of 45?s at 94?��C, 1?min at 58?��C, and 2?min at 72?��C, followed by a final extension step at 72?��C for 10?min. Amplification of archaeal 16S rRNA genes was tested for replicates B of enrichment http://www.selleck.cn/products/pfi-2.html II using the primers arch344f (5�� ACG GGG TGC AGG CGC GA 3��) and arch915R (5�� GTG CTC CCC CGC CAA TTC CT 3��) following the protocol of Karr et?al. (2006). PCR products http://www.selleckchem.com/products/MG132.html were purified using the Qiagen MinElute PCR Purification kit (Medicorp, Inc., Montr��al, QC, Canada). For T-RFLP analysis, purified PCR products were checked and quantified by agarose gel electrophoresis applying the E-Gel? Low Range quantitative DNA ladder (Invitrogen, Carlsbad, CA). Approximately 80?ng of DNA was digested using the restriction enzyme MspI according to the manufacturer's protocol (Fermentas Canada Inc., Burlington, ON, Canada), and the DNA fragments were purified by ethanol precipitation. Depending on the source of the terminal restriction fragments (T-RFs), 10�C30?ng (clones) or 30�C60?ng (enrichment cultures) was loaded on the capillary sequencer CEQ? 8000 Genetic Analysis System (Beckman Coulter Canada Inc., Mississauga, ON, Canada). GenomeLab DNA Size Standard kit �C 600 was used as internal standard. Fragment lengths issued by the analysis software (CEQ 8000 Genetic Analysis System version 9.0.25 Fragment Version 2.3.4) were rounded to whole numbers of nucleotides (nt). Fragments with a difference in lengths of less than two nucleotides were considered identical. http://www.selleckchem.com/products/epacadostat-incb024360.html Relative abundances of fragments were expressed as percentage of total peak area. Fragments with a peak area