"Urine-fed Control, reactor A-3, sampled 9/2/13"
Oligonucleotide primers targeting the 16S rDNA gene were selected according to Klindworth et al.  The nucleotide sequences of the primers were 5’-TCGTCGGCAGCGTCAGATGT GTATAAGAGACAGCCTACGGGNGGCW GCAG-3’ (forward) and 5’-GTCTCGTGG GCTCGGAGATGTGTATAAGAGACAGG ACTACHVGGGTATCTAATCC-3’ (reverse), and expected to amplify a ~550 base pair section of the DNA target (Illumina, 2013; Klindworth et al., 2013).  Reaction mixtures were prepared in accordance with Illumina’s recommendations in a total volume of 25 µL, using 2.5 µL of extracted DNA at 5 ng/µL, with 5 µL of each 1 µM forward and reverse primers and 12.5 µL of 2x KAPA HiFi HotStart ReadyMix. Thermal cycling was carried out with an initial denaturation step of 98°C for 20 s, as recommended by the KAPA HotStart manufacturer, followed by a second denaturation step at 95°C for 3 minutes.  Amplification was performed for 25 cycles of denaturation at 95°C for 30 s, annealing at 55°C for 30 s, and elongation at 72°C for 30 s, then completed by a final elongation step of 72°C for 5 min (Illumina, 2013). 
9-2A3-1