GSM9787223: mdx mice treated with high dose PB1046-3; Mus musculus; RNA-Seq
Frozen cell pellets (isolated cardiomyocytes) were thawed at room temperature, and total RNA was extracted using the RNeasy Fibrous Tissue Mini Kit (74704, QIAGEN) according to the manufacturer's instructions. Samples were run on the Agilent Tapestation 4200 to determine level of degradation thus ensuring only high quality was used (RIN Score>=8). The Qubit fluorimeter was used to determine the concentration prior to starting library prep. 1.0 μg of total RNA (DNAse treated) was then prepared with the TruSeq Stranded mRNA Library Prep Kit from Illumina. Poly-A RNA was purified and fragmented before strand specific cDNA synthesis. cDNA were then a-tailed and indexed adapters were ligated. After adapter ligation, samples were PCR amplified and purified with AmpureXP beads, then validated again.