16S amplicon sequencing of chicken gut metagenome: Vaccination Status - Mock-Vaccinated; Days Post Inoculation - D7; Replicate - R.6
Total microbial DNA was extracted from chicken cecal content samples using the QIAamp PowerFecal Pro DNA Kit. DNA was assessed qualitatively by agarose gel electrophoresis and quantitatively using the QubitTM dsDNA Broad Range Quantitation Kit. DNA samples were standardized to 10 ng/L in UltraPure distilled water, and PCR amplification of the V4 region of 16S rRNA gene was performed as previously described. Successful amplification was confirmed by agarose gel electrophoresis and QubitTM dsDNA High Sensitivity Quantitation Kit. Individual samples were normalized using SequelPrepTM Normalization Plates, pooled, and concentrated using the Zymo Research DNA Clean and Concentrator.