GSM7140777: EarSkin_MC903_IL26_IgG_1; Mus musculus; RNA-Seq
For RNA isolation, half a mouse ear was incubated in 1 mL RNAprotect (Qiagen, #76104) for 24 hours at 4 °C.  Subsequently, skin tissue was homogenized in 500 μL RLT RNA lysis buffer (Qiagen, #79216) containing 40mM dithiothreitol (DTT, Sigma, #3483-12-3) using tissue homogenizing CK14 tubes (bertin technologies, #KT03961-1-203.0.5) and a PrecellysTM Evolution SUPER Homogenizer (bertin technologies) for 2 cycles with 3x20 s at 6400 rpm at 4 °C and RNA was prepared using the RNeasy Mini kit (Qiagen, #74106) according to manufacturer's instructions. After tissue disintegration RNA was isolated using the RNeasy® Mini Kit (Qiagen, #74106) in combination with on column DNA digestion according to manufacturer's instructions. TruSeq libraries were prepared from 200 ng of total RNA using the TruSeq Stranded Total RNA Library Prep Gold kit (Illumina, #20020599), according to the manufacturer's instructions. Ribo-Zero