title: GSM9040796: PBS_PBS3; Mus musculus; RNA-Seq
library_construction_protocol: After mice anesthesia, the thoracic cavity was dissected, lungs were removed after PBS perfusion via the heart and subsequently placed in a 15 mL test tube containing 5 mL RNAlater (Millipore Sigma). Lungs were weighted and homogenized in RNAlater until smooth using bead beater. RNA was isolated from lungs using Qiagen RNeasy mini kits (Qiagen, CA, USA) according to the manufacturers' instructions, then submitted for RNA-sequencing at LC sciences (Houston, Texas) Poly(A) RNA sequencing library was prepared following Illumina's TruSeq-stranded-mRNA sample preparation protocol. RNA integrity was checked with Agilent Technologies 2100 Bioanalyzer. Poly(A) tail-containing mRNAs were purified using oligo-(dT) magnetic beads with two rounds of purification. After purification, poly(A) RNA was fragmented using divalent cation buffer in elevated temperature. The DNA library construction is shown in the following workflow. Quality control analysis and quantification of the sequencing library were performed using Agilent Technologies 2100 Bioanalyzer High Sensitivity DNA Chip. Paired-ended sequencing was performed on Illumina's NovaSeq 6000 sequencing system.