title: Effect of glucose on nitrogen removal and microbial community in anammox-denitrification system
design: DNA was extracted using the Powersoil DNA isolation kit (MiBio Laboratories, Inc., USA) according to the manufacturer’s instruction. 16S rRNA genes segments were PCR-amplified using forward primer 515F (5’-GTGCCAGCMGCCGCGGTAA-3’) and reverse primer 806R (5’ -GGACTACHVGGGTWTCTAAT-3’) for the V4 region. The PCR reaction mixture contained 10 × Ex Taq Buffer (6 uL), dNTP (6 uL), BSA (0.6 uL), Ex Tap (0.3 uL), Primer F (1.2 uL), Primer R (1.2 uL), DNA (1 uL) and ddH2O (43.7 uL).The PCR thermal program was performed at 94 ℃ for 5 min; 31 cycles of 94℃ for 30 s, 52 ℃ for 30 s, and 72 ℃ for 45 s; and a final extension at 72 ℃ for 10 min.