MiSeq_wat_microbial_Ebro_river_delta
For each sample, Illumina compatible, dual indexed amplicon libraries of the 16S-V4 rRNA hypervariable region were created with primers 515f/806r using the method developed by the Schloss. Completed libraries were batch normalized using Invitrogen SequalPrep DNA Normalization Plates; product recovered from the plats was pooled. The pool was QCd and quantified using a combination of Qubit dsDNA HS, Agilent 4200 TapeStation High Sensitivity DNA and Kapa Illumina Library Quantification qPCR assays. This pool was loaded a standard Illumina MiSeq v2 flow cell and sequencing was performed in a 2x250bp paired end format using a MiSeq v2 500 cycle reagent cartridge. Custom sequencing and index primers complementary to the 515/806 target sequences were added to appropriate wells of reagent cartridge. Base calling was done by Illumina Real Time Analysis (RTA) v1.18.54 and output of RTA was demultiplexed and converted to FastQ format with Illumina Bcl2fastq v2.19.1. A summary of the run output is attached below. Basic QC information about your sequence data is provided by the accompanying FastQC reports.