title: Shotgun sequencing of wheat root associated bacteria
design: DNA extraction in triplicates from the same batch was combined to get three replicates per treatment and 12 sample all together. Shotgun metagenome libraries were prepared using the Celero DNA-seq library preparation kit (NuGen, Takara Bio, USA) with enzymatic shearing, according to the manufacturers instructions. Subsequently, all libraries were pooled in equal volume and size selection (350-400 bp fragments) was performed using a Blue PippinPrep instrument (Sage Scientific). The libraries were then sequenced using an Illumina MiniSeq instrument employing a mid-output kit. Based on the number of reads per sample, the samples were re-pooled with varying volumes, and size selection was performed again using the same size range. The final size-selected pool was sequenced on an Illumina NovaSeq instrument with an S4 flow cell, employing 2x150 base reads. Library preparation and pooling was performed at the University of Illinois at Chicago Sequencing Core (UICSQC), and sequencing was performed by Novogene Corporation (Chula Vista, CA).
library_name: 400ppm_CO2_100ppm_Nitrate_biological_replicate_2