16S rRNA reads of anaerobic sludge fed with palm oil mill effluent: without supplement
Sequencing of the 16S rRNA amplicon was conducted using the Illumina MiSeq platform with a paired end 2x250 bp read length. The DNA of the samples was extracted using the DNeasy PowerSoil Pro DNA Kit (Qiagen, USA) and amplified with 341F and 805R as the forward and reverse primers, respectively (targeting the V3-V4 variable regions). The DNA library was prepared with 2X sparQ HiFi PCR Master Mix (QuantaBio, USA), and amplification was performed under the following conditions: 2 min initial denaturation at 98 C, followed by 30 cycles of 98 C for 20 s, 60 C for 30 s, and 72 C for 1 min, and a single final extension step at 72 C for 1 min. The amplicons obtained were purified with sparQ Puremag Beads (QuantaBio, USA) and later indexed with 5 l of Nextera XT index primers in a 50 l PCR. Subsequently, 8-10 cycles of PCR were run under the abovementioned conditions.