GSM8387257: PBS Group; Mus musculus; RNA-Seq
Total RNA was extracted from NLF using Trizol, and the RNA integrity and quantity were assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system. Upon confirming the RNA quality standards (total RNA ≥ 500ng, RNA integrity number ≥ 5.8), the RNA library for meta-transcriptome sequencing was prepared by depleting ribosomal RNA using an rRNA Removal Kit, fragmenting the RNA into 250-300 bp pieces, and reverse-transcribing these into cDNA. Subsequent steps included end repair, adenylation, and adaptor ligation. The library was then size-selected, amplified, and its concentration and insert size were validated using Qubit® fluorometer, Agilent 2100 Bioanalyzer, and qPCR before sequencing.