GSM8701778: Wildtype_1; Mus musculus; RNA-Seq
TA muscles were dissected at 22 months of age, followed by total RNA isolation using an RNA isolation kit (Qiagen, cat# 763134), according to the manufacturer's instructions. Total RNA concentration and quality (A260/280 ratio ≥2.0) were evaluated using a Nanodrop spectrophotometer (ThermoFisher Scientific). The quality of total RNA was further measured using the Agilent 2100 Bioanalyzer (Agilent, Santa Clara, CA). After Sample QC was by qubit and Agilent Bioanalyzer, a certain amount of RNA samples were denatured at suitable temperatures to open-up their secondary structure, and mRNA is enriched by oligo (dT) -attached magnetic beads followed by mRNA fragmentation and cDNA Synthesis. The double-stranded cDNA fragments were subjected to end-repair, the addition of a single 'A' nucleotide to the 3' ends of the blunt fragments and ligated to adaptors before amplification. After quality control of the prepared library, the amplified products were denatured to generate single stranded PCR products which was then circularized to replicate using rolling cycle amplification yielding a DNA nanoball (DNB) containing multiple copies of DNA is generated. Sufficient quality DNBs are then loaded into patterned nanoarrays using high-intensity DNA nanochip technique and sequenced through combinatorial Probe-Anchor Synthesis (cPAS).