GSM7916033: Mst1/2 KO sample 1; Mus musculus; RNA-Seq
The esophagus was dissected from the mice and muscle layers and adventitia were stripped away using forceps. Subsequently, the esophagus was incubated with 0.5 U/ml of Dispase I at room temperature for 10 minutes, allowing the separation of mesenchymal layers from the epithelium. The mesenchyme layers were then removed from the epithelium under a dissection microscope, and the epithelium was then washed with sterile 1X PBS and lysed with RNA lysis buffer. Total RNA was extracted from the lysate using the Qiagen RNeasy Plus Universal Mini Kit. Libraries were constructed using the VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina (Vazyme Biotech) and size-selected with VAHTS DNA Clean Beads (Vazyme Biotech) according to the manufacturer's instructions. The libraries were sequenced by Illumina NovaSeq 6000 using a paired-end (150 bp for each end of the fragment) configuration.