Floral Nectar Asthens 2
Floral nectar DNA samples was amplified by triplicates 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). PCR products were pooled before the second amplification. 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.