title: Marine Microbial Biofilms-Bac
design: The universal 16S Eubacterial primers 27F-519Rmodified were used in a single-step 30 cycle PCR using HotStarTaq Plus Master Mix Kit (Qiagen, Valencia, CA) and under the following conditions: 94oC for 3 minutes, followed by 28 cycles of 94oC for 30 seconds;  53oC for 40 seconds and 72oC for 1 minute. A final elongation step at 72oC for 5 minutes was performed.  Subsequently, all amplicon products from all samples were mixed in equal concentrations and purified using Agencourt Ampure beads (Agencourt Bioscience Corporation, MA, USA). Sequencing was performed at MR DNA (www.mrdnalab.com, Shallowater, TX, USA) utilizing Roche 454 FLX titanium instruments and reagents and following the manufacturer’s guidelines.