title: RNA-Seq of biofilm in stream  Ecker sample num_b14
design: Subsample for microbial diversity were frozen at 75 C once in the laboratory. DNA was extracted using the DNeasyBlood & Tissue Kit (Qiagen). After extraction, samples exhibiting a brown color  underwent an additional purification and concentration step using Amicon Ultra centrifugal filters. DNA yield was measured with either a Qubit fluorometer or a NanoDrop photometer. Samples < 5ng/l were discarded from further analyses. The remaining samples were subjected to two consecutive PCR amplifications. The library was sequenced on the Illumina MiSeq platform using the MiSeq Reagent Kit v3 (2 x 300 bp) The primer sequence for 16S rRNA gene were 16S_Illu_F (tcgtcggcagcgtcagatgtgtataagagacagcctacgggnggcwgcag) and 16S_Illu_R (gtctcgtgggctcggagatgtgtataagagacaggactachvgggtatctaatcc).