GSM4500890: S9; Mus musculus; RNA-Seq
Total RNA was isolated from day 10 snap frozen wound samples using the Qiagen miRNEasy kit (Qiagen, 217004) on a TissueLyser LT (Qiagen) per the manufacturer's instructions. RNA samples were quantified using Nanodrop 2000 spectrophotometer (ThermoFisher) and Qubit 2.0 Fluorometer (Life Technologies) and RNA integrity was checked with 4200 TapeStation (Agilent Technologies). rRNA depletion was performed using Ribozero rRNA Removal Kit (Illumina).  RNA sequencing library preparation used NEBNext Ultra RNA Library Prep Kit for Illumina by following the manufacturer's recommendations (NEB). Briefly, enriched RNAs were fragmented for 15 minutes at 94 °C. First strand and second strand cDNA were subsequently synthesized.  cDNA fragments were end repaired and adenylated at 3'ends, and universal adapter was ligated to cDNA fragments, followed by index addition and library enrichment with limited cycle PCR. Sequencing libraries were validated using the Agilent Tapestation 4200 and quantified by using Qubit 2.0 Fluorometer as well as by quantitative PCR (Applied Biosystems).