amplicon sequencing of the prokaryotic V4 region (23S rRNA): Koutsourelia.Spring
Amplicon sequencing analysis by targeting the V4 region of the 16S rRNA gene for prokaryotes (amplified with 515f - 806r primer pair). A two-step amplification protocol was implemented, to construct amplicon libraries. A unique 12-bp index was added to the forward primer that served as a barcode to identify sequencing products in multiplexed libraries. 2x250 bp pair-end sequencing via the HiSeq2500 platform (Illumina, San Diego, USA) was performed at the Genome sequencing centre of the Brigham Young University (Provo, UT, USA).