Nodule Viablecells Rep 3
We used a DNA dye, SYBR to determine viable microbial cells from soil particles and debris with flow cytometry. Viable cells were sorted with FACS. We exctracted DNA from pelleted cells with prepGEM Bacteria Kit cell lysing kit following manufacturer instructions (New England Biolabs) .  We used 515f-Y (GTGYCAGCMGCCGCGGTAA, Parada 2016), and 806R-B GGACTACNVGGGTWTCTAAT, Apprill 2015) primers to amplify the 16S rRNA gene with illumina adapter overhangs. Samples sequenced on Illumina MiSeq platform using 250 bp x 250 bp paired-end sequencing and aiming at 100,000 reads per sample