16s RNA from  Rust at ascidian/steel interface
DNA extracted from sediment and rust were used for sequencing. The integrity and approximate length of the DNA were examined by 1% agarose gel electrophoresis. For Illumina HiSeq sequencing, 16S rRNA genes from bacteria and eukarya were amplified using the primer sets 515F/806R and 528F/706R, respectively. PCR products were purified with a PCR purification kit (QIAGEN, Hilden, NRW, Germany). The PCR libraries were conducted using TruSeq DNA PCRFree specimen Preparation Kit (Illumina, San Diego,CA). After quantification with Qubit, the PCR libraries were sequenced on the Illumina NovaSeq6000 platform.