Symbiodiniaceae ITS2 rDNA metabarcoding
Theregionoftheinternaltranscribedspacer2(ITS2)nrDNA(~400bp)wasamplifiedbyPCRwiththespecificprimersSYM_VAR_5.8S2andSYM_VAR_REV(Humeetal.,2018)linkedtoIlluminaadaptersfollowingthe16SMetagenomicSequencingLibraryPreparationIlluminaprotocol.AmplificationsintriplicatewerepooledandPCRclean-upwasperformedwiththeProNexSystem.Illuminaindexingprimers(NexteraXTIndexKit96indexes,384samples)wereaddedto5LofmixedpurifiedPCRproducts,accordingtothemanufacturerinstructions.Theproductspooledinequimolaramounts(4nM)andsequencedontheIlluminaMiSeqplatformusingaMiSeqReagentKitv2(500-cycles)paired-endrunwith25%PhiX.