GSM7440695: soleus, Female, B6, HF, Age 5 Week, Replicate 4; Mus musculus; RNA-Seq
Total RNA was isolated from the tissue using RNeasy Plus Universal Kit (Qiagen, Valencia, CA, USA) according to the manufacturer's instructions. All RNA samples were assessed using the Bioanalyzer 2100 and quantified using the Thermo Fisher Qubit RNA assay. Each RNA sample with a RIN value above 8 and a quantity of 1000ng was utilized to generate a Poly(A)-based RNAseq library following the Illumina TruSeq stranded mRNA protocol. Briefly, the poly(A) RNAs were purified using oligo dT magnetic beads and were then fragmented with divalent cations, followed by reverse transcription into cDNA. The Illumina paired-end oligo adapters were ligated to the cDNA fragments, and the RNAseq libraries were amplified using PCR. The finished libraries were purified and analyzed on an Agilent Bioanalyzer 2100, revealing an average library size ranging from 285 to 300 base pairs. To determine the quantity of these libraries, they were further measured using the Thermo Fisher Qubit HS dsDNA assay.