MetNeo_metagenome_day4_1
DNA quality control and dilution. The DNA was isolated and quantified by using Quant-iT 1X dsDNA Assay Kits, BR (Life Technologies, #Q33267) in combination with Varioskan LUX Microplate Reader (Thermo Fisher Scientific, #VL0000D0). The DNA was then diluted in water for the following library preparation. Library preparation and sequencing. The sequencing libraries were prepared with the Illumina DNA Prep, (M) Tagmentation (96 Samples, IPB) kit (Illumina, #20060059) in combination with the UD for Illumina DNA/RNA UD Indexes (Illumina, #2009167/8/9) and the amplified libraries were purified with the double-sided bead purification procedure, as described by the Illumina protocol. Then, libraries concentration (ng/l) were quantified with the Quant-iT 1X dsDNA Assay Kits, HS (Life Technologies, #Q33232) in combination with the Varioskan LUX Microplate Reader (Thermo Fisher Scientific, #VL0000D0). In addition, the base pair length (bp) was evaluated by using the D5000 ScreenTape Assay (Agilent Technologies, #5067-5588/9) in combination with the TapeStation 4150 (Agilent Technologies, #G2992AA). By knowing both library concentration and base pair length, it is possible to obtain the correct library volume to pool in the same tube in order to achieve optimal cluster density. The library pool was then quantified with the Qubit 1x dsDNA HS kit (Life Technologies, #Q33231) through the Qubit 3.0 Fluorometer (Life Technologies, #Q33216) and the base pair length (bp) was evaluated as described before. Finally, the library pools were sequenced using the Novaseq X Plus platform (Illumina) at an average depth of 7.5 Gb per sample.