Metagenomics and metabarcoding experimental choices and their impact on microbial community characterization in freshwater recirculating aquaculture systems

Table 1 Primers used in this study

Targeted region	Short-hand	FW primer	FW primer sequence	FW primer length	FW melting temperatures	REV primer	REV primer sequence	REV primer length	REV melting temperatures	Amplicon length	Primer target species
16S: V3-4	MiSeq	341F	5′-CCT ACG GGN GGC WGC AG-3′	17 bp	58–60 °C	805R	5′-GAC TAC HVG GGT ATC TAA TCC-3′	21 bp	52–60 °C	~ 465 bp	Bacteria
16S: V4	Earth	515F	5′-GTG YCA GCM GCC GCG GTA A-3′	19 bp	64–68 °C	806R	5′-GGA CTA CNV GGG TWT CTA AT-3′	20 bp	54–58 °C	~ 300 bp	Bacteria & Archaea
16S: V1-3	–	27F	5′-AGA GTT TGA TCC TGG CTC AG-3′	20 bp	58 °C	534R	5′-ATT ACC GCG GCT GCT GG-3′	17 bp	56 °C	~ 500 bp	Bacteria
16S-ITS-23S	PacBio	–	Proprietory	–	–	–	Proprietory	–	–	2500 bp	Bacteria

Illumina MiSeq forward and reverse primers carried overhang adapters (5′-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-Forward primer, 5′-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-Reverse primer) for compatibility with Illumina index and sequencing adapters
FW Forward primer, REV Reverse primer, bp Base pairs

ISSN: 2524-6372