4.2. DNA Extraction, PCR, and Amplicon Sequencing
BLOOD, STERILE
DNA was extracted from the eNAT tubes using the Qiagen Blood Mini Kit with several modifications. Briefly, 1 mL of sample was aliquoted in sterile 2 mL tubes containing 1.0 mm and 0.1 mm sized silicon beads. The tube was centrifuged at 10,000× The 16S rRNA gene V4 region was amplified using 515F and 806R primers with 50 ng of DNA in final volume of 50 µL. Several negative controls were amplified using water as a template. The amplification and amplicon size were confirmed by running samples on 1% (