GSM8758923: NaCl - 12 weeks - 2; Mus musculus; RNA-Seq
Lung lobes were stored in RNA later solution (Invitrogen) to prevent cellular RNA degradation during tissue thawing and to protect its integrity. Samples were disrupted using TissueRuptor II (Qiagen) in RNA Lysis Buffer (RLT, Qiagen) supplemented with 10% b-mercaptoethanol. Total lung RNA was isolated using RNeasy Mini Kit (Qiagen) according to the manufacturer's protocol. Only the RNA with high integrity (RIN>8) was subjected to cDNA library preparation according to TruSeq Stranded Total RNA protocol (Illumina), followed by the quality confirmation (Agilent Technologies 2100 Bioanalyzer). cDNA libraries were prepared for sequencing using standard Illumina protocols.