title: 16S sequences of EUPHYLLIA PARADIVISA microbiome: tissue
design: Bacterial 16S rRNA genes were amplified using 27F and 1100R primers and then a nested PCR was performed using 27F and (Hongoh et al., 2003) and  338R (Fierer et al., 2008) primers. PCR mixtures (25  l) were prepared in duplicate and each contained 2  l of DNA template, 5  l of 10_PCR buffer (50mM KCl, 10mM Tris-HCl and 1.5mM MgCl2), 200 mM of dNTP, 0.2 mM of each primer and 2.5U Taq polymerase (Promega, Madison, WI, USA). The PCR thermal regime consisted of an initial denaturation of 3 min at 98  C, followed by 20 cycles of 30 s at 98  C, 30 s at 57  C, 1min at 72  C and a final cycle of 5 min at 72  C. The tagged amplicons NGS was performed on an IonTorrent (Applied Biosystems) at the Bar-Ilan University sequencing unit