GSM4904946: ND_PMA_1; Mus musculus; RNA-Seq
Mouse ear tissues were homogenized in Trizol using a bead cells disrupter/micro homogenizing system (Tomy Micro Smash MS-100). Total RNA was isolated following the double extraction protocol: RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction (TRIzol, Thermo Fisher Scientific) followed by a Qiagen RNeasy Micro clean-up procedure according to the manufacturer's protocol. cDNA libraries were prepared from 2 ng total RNA starting material and 1 ul of a 1:50,000 dilution of ERCC RNA Spike in Controls (Ambion® Thermo Fisher Scientific) using the SMARTSeq v2 protocol with the following modifications: 1. Addition of 20 µM TSO; 2. Use of 250 pg cDNA with 1/5 reaction of Illumina Nextera XT kit (Illumina, San Diego, CA, USA). The length distribution of the cDNA libraries was monitored using a DNA High Sensitivity Reagent Kit on the Perkin Elmer Labchip (Perkin Elmer, Waltham, MA, USA). All samples were subjected to an indexed paired-end sequencing run of 2x151 cycles on an Illumina HiSeq 4000 system (Illumina) (32 samples/lane).