Production of Scenedesmus sp. in pilot-scale thin-layer cascade reactors: Influence of water type on biomass composition
DNA from the freeze-dried biomass samples (av. 95 mg) was extracted in triplicate for each sample using the DNeasy Plant Kit (Qiagen, Germany) according to manufacturer instructions. The yield and purity of the extracted DNA was quantified on a Nanodrop 1000 spectrophotometer (Thermo Fisher Scientific) while eventual fragmentation was determined through gel electrophoresis 1% (w/v) 1TAE agarose gels. Illumina sequencing was performed on all samples for prokaryotic and eukaryotic communities at Stab Vida Lda (Lisbon, Portugal). For bacteria, the V3-V4 hypervariable region on the 16S gene was selected and amplified with the primers Bakt_341F: CCTACGGGNGGCWGCAG and Bakt_805R: GACTACHVGGGTATCTAATCC (Herlemann et al., 2011) while for eukaryotes the 18S V9 region was amplified with the primers 1389F: TTGTACACACCGCCC, 1510R: CCTTCYGCAGGTTCACCTAC was targeted (Amaral-Zettler et al. 2009). The generated DNA libraries were sequenced with MiSeq Reagent Kit Nano in the lllumina MiSeq platform, using 300bp paired-end sequencing reads.