title: 16s rDNA metagenomic sequence analysis of bacterial community isolated from the anodic chamber of microbial fuel cell fed with lipid extracted algae biomass.
design: The genomic DNA of mixed bacterial culture prevailing on anode electrode in a microbial fuel cell, was isolated by using Soil DNA purification kit, Himedia. Illumina plate based Next Generation Sequencing (NGS) method was used to sequence the V3-V4 hyper-variable region of 16s rDNA at Xcleris genomics facilty (Ahmedabad, India), to analyze the diversity of anodic microbial community growing on lipid extracted algae (LEA) biomass. The V3-V4 hyper-variable region of bacteria and archaea was amplified using the primer pair (Prokaryote-V3 forward:5CCTACGGGNBGCASCAG3 and Prokaryote-V4-reverse-5 GACTACNVGGGTATCTAATCC3). The amplicon libraries were constructed using the amplified V3-V4 hyper-variable region. The data obtained from NGS platform was analyzed using QIIME (Quantitative Insight Into Microbial Ecology) tool. UCLUST algorithm was utilized to assign an operational taxonomic unit (OTU) which is a representative taxonomic unit of similar sequence pooled together on the basis of 97% sequence resemblance. Green gene database was employed to make taxonomic assignments at 90% threshold value.