RNA-Seq of Surface Sea Water: Pari Island
Seawater samples were collected from Ascidia sp. In-situ environment in Pari Islands. Water samples were aseptically filtered using a vacuum filter with a 0.22-micron nitrocellulose filter paper. The filter paper used for filtration was then employed in the subsequent process. DNA extraction was performed using the ZymoBIOMICS DNA Kit (Irvine, California, USA). The extracted DNA was then quality-controlled (QC) using agarose gel electrophoresis and the Qubit BR-DNA Assay. Shotgun metagenomic was carried out using MGI DNBseq50 (Wuhan, China). DNA concentration was evaluated using the Qubit fluorometer dsDNA BR Assay with the condition that the DNA concentration value for sequencing was 15 ng/L. The DNA library was fabricated through end-repaired adapter and polymerase chain reaction amplification using the MGIEasy FS DNA Library Prep Kit (Wuhan, China). The library then transformed the single-strand circularized DNA library into DNA nanoballs (DNB) and sequenced by DNBSeq-G50 with an average read length of equal 150bp. The metagenomic assembly was performed using Metaspades. The process of genome assembly from metagenomic samples generates scaffolds. The results of the metagenomic assembly are evaluated and compared with the closest reference using metaQUAST. Functional and taxonomic analysis were performed using Diamond + MEGAN. Diamond was used to align all DNA fragments to a protein database. The alignment results were then analyzed for taxonomic and functional information using MEGAN.