RNA-seq of Sapata outside cave replicate 1
The 16S rRNA gene V4 variable region PCR primers 515/806 with barcode on the forward primer were used in a 30 cycle PCR under the following conditions: 94 C for 45 sec, annealing at 55 C for 1 min, extension at 72 C for 1 min and a final extension at 72 C for 10 min. Amplification products were purified using Agencourt AMPure XP kit (Beckman Coulter) and quantified by real-time PCR, according to the manufacturers protocol Kit KAPA Library Quantification Kit - Illumina/Universal (Roche). An equimolar pool of DNA was generated by normalizing all samples at 4 nM for the sequencing, which was conducted using the Illumina MiSeq new generation sequencing system (Illumina Sequencing) and two units of the MiSeq Reagent V2 Micro 300 cycles kit  2 x 150 bp reading with 20% PhiX Sequencing Control V3 (Illumina).