title: acdS gene amplicon sequencing of bulk or rhizosphere soil
design: Partial acdS gene was amplified in duplicate using the primers acdSF5 (GGCAACAAGMYSCGCAAGCT) and acdSR8 (CTGCACSAGSACGCACTTCA). Primers were equipped with Illumina Nextera XT Index Kit adapter sequences. PCR was performed using Kapa Hifi HotStart ReadyMix with the cycling conditions: 30 cycles of 65C for 10 s, 72C for 10 s. PCR products were purified and an index PCR was performed using the Illumina Nextera XT Index Kit and Kapa Hifi HotStart ReadyMix. The index PCR products were purified,  quantified, pooled equimolarly and sequenced on an Illumina MiSeq system to 2 x 150 nucleotide lengths.