16s RNA V4 amplicon for human in atherosclerosis
Collected stool samples were stored at -80C until DNA isolation. Total DNA was extracted from 250 mg of the whole fecal samples using the QIAamp Fast DNA stool kit (QIAGEN). The hypervariable V4 region of the 16s rRNA gene was amplified by polymerase chain reaction (PCR) with 515-806 primers tailed with sequences to incorporate Illumina flow cell adapters and indexing barcodes. Sequencing was carried out on an Illumina Miseq platform (2  250).