GSM5493528: Lung, GP_D3_3B; Mus musculus; RNA-Seq
Mouse lungs were homogenized in liquid nitrogen. 1 ml Trizol was added per 50-100 mg of tissue and the homogenate was incubated at room temperature for 5 minutes. 200 μl chloroform were added per ml of Trizol, shaken vigorously for 15-30 seconds and incubated at room temperature for 15 minutes. Samples were centrifuged for 15 minutes at 4˚C and 12,000 rcf. The aqueous phase was carefully collected in a separate tube. 500 μl isopropanol was added per 1 ml Trizol, inverted to mix and incubated at room temperature for 5-10 minutes. Samples were transferred to Qiagen RNeasy Spin Columns and RNA was isolated according to the manufacturer's instructions. RNA-seq libraries were constructed using the Illumina TruSeq Stranded Total RNA Library Prep Kit.