Stool healthy IgA : HC-012
Fresh stool samples were resuspended at 100 g/mL, filtered for 30 m and stored in glycerol medium at -80 C. Frozen stocks were thawed and stained with anti-human IgA1 and anti-human IgA2 antibodies. Sorting of IgA bound events was done using at a BD Influx cell sorter. The sorted bacteria were washed and directly subjected to the Illumina Nextera XT workflow using the primers 5-TCgTCggCAgCgTCAgATgTgTATAAgAgACAgCCTACgggNggCWgCAg-3 and 5-gTCTCgTgggCTCggAgATgTgTATAAgAgACAggACTACHVgggTATCTAATCC-3 for the first amplicon PCR. After Nextera v2 index PCR and quality control by capillary gel electrophoresis samples were pooled and sequenced on MiSeq device (Illumina).