RNA-seq of Cymodocea nodosa: Rhizome
The initial lysing step for the isolation kit (Qiagen) required both a lysing solution and mechanical bead beating for 10 min, we extended this to 1 hr 10 min in order to maximize DNA yields. Extracted DNA was stored at -20C until 16S rRNA amplicon sequencing. Bacterial communities of each sample were analyzed using 454 pyrosequencing of the 16S rRNA gene. Samples were sequenced by MRDNA (www.mrdnalab.com , MRDNA, Shallowater, TX) using a single-step 30 cycle PCR, using primers specific for the 16S gene (27F and 519R). Samples were then sequenced using a Roche 454 FLX titanium sequencer as per the manufacturers guidelines.