Water F Autumn 1st year
"Water DNA samples was amplified using primers targeting the V4 variable region of the bacterial 16SrRNA gene. Primers sequences were: CS1_515F (ACACTGACGACATGGTTCTACAGTGCCAGCMGCCGCGGTAA) and CS2_806R (TACGGTAGCAGAGACTTGGTCTGGACTACHVGGGTWTCTAAT) (Caporaso et al. 2012). These primers contained 5' common sequence tags, in accordance to Moonsamy et al. (2013). The PCR amplification procedure is described in details in Aizenberg-Gershtein et al. (2015). Illumina MiSeq sequencing. MiSeq sequencing was performed at the DNA Services (DNAS) Facility - University of Illinois, Chicago (UIC). The sequencing protocol is described in Aizenberg-Gershtein et al. (2015). The procedure included a second PCR amplification in 96-well plates, where each well received a separate primer pair, obtained from the Access Array Barcode Library for Illumina Sequences [10-base barcode (Fluidigm, South San Francisco, CA; Item no. 100-4876)]. Pooled, diluted libraries were sequenced using a MiSeq 600-cycle sequencing kit version 3, and analyzed with Casava1.8 (pipeline 1.8). Reads were 200 nucleotides in length (paired end, 2x200). PhiX DNA was used as a spike-in control. Barcode sequences were provided to the MiSeq server, and were automatically binned according to 10-base multiplex identifier (MID) sequences. Raw reads were recovered as FASTQ files."