title: GSM5777285: Colon flush, Naive3; Mus musculus; ncRNA-Seq
library_construction_protocol: colon:  Gut contents were collected by conducting a colon flush, Bacterial genomic DNA was isolated, lung:  Quick-DNA Fungal/Bacterial Midiprep Kit was used in the isolation of bacterial genomic DNAs from lungs. Briefly, 500 mg of lung tissue was added to ZR Bashing/Bead lysis filtration tube with 6 ml of genomic lysis buffer containing β-mercaptoethanol (0.5% v/v). The tubes were placed in a 50ml–tube holder for 1 minute, followed by centrifugation at 3,000 g for 5 minutes. Following centrifugation, the samples were filtered through Zymo-Spin V-E columns in a vacuum manifold with a vacuum pressure of 600 mmHg. After filtering, the columns were transferred to collection tubes for centrifugation at 10,000 g for 1 minute. The columns were washed with 400 µl of genomic DNA-wash buffer and then DNA samples were eluted from the columns with 150 µl DNA Elution Buffer under centrifugation at 10,000 g for 1 minute. DNA samples were stored at -80ºC and used in the library preparation for 16S ribosomal RNA sequencing using 16S Metagenomic Sequencing Library Preparation Kit ribosomal RNA sequencing