non IgE allergy pediatric cohort microbiome - Romania
Description
Pediatric cohort -non IgE food allergy - 16S rDNA amplicon sequencing; cohort characteristics metadata
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Steps to reproduce
Bacterial community profiling was performed by amplification of the V3–V4 hypervaria-ble regions of the 16S rRNA gene using universal primers compatible with Illumina over-hang adapters (341F/806R). PCR amplification was conducted under optimized cycling conditions, and amplicons were purified using magnetic bead-based cleanup. Indexed sequencing adapters were added in a second PCR step, followed by purification and quantification. Libraries were pooled at equimolar concentrations and sequenced on an Illumina MiSeq platform (Illumina, San Diego, CA, USA) using paired-end 2 × 30250 bp chemistry. Negative extraction controls and no-template PCR controls were included to monitor contamination. Sequencing quality was assessed using standard Illumina metrics