16S rRNA gene of Peridinium free-living bacteria: dispersed oil
The V4 hyper-variable region of the bacterial 16S rRNA gene was amplified using primers 515f/806r. Completed libraries were normalized using Invitrogen SequalPrep DNA Normalization plates and normalized products were pooled. The pooled PCR products were cleaned up using AMPure XP magnetic beads. The completed library pool was QCÕd and quantified using a combination of Qubit dsDNA HS, Caliper LabChipGX HS DNA and Kapa Illumina Library Quantification qPCR assays. The amplicon pool was loaded on an Illumina MiSeq v2 Standard flow cell and sequencing was performed in a 2x250bp paired end format using a v2 500 cycle MiSeq reagent cartridge. Custom sequencing and index primers were added to appropriate wells of the 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 v2.19.0.
SUB3153066
P-FL-OD3