RNA-Seq of Human LHCN-M2 skeletal muscle myotubes to Electrical Pulse Stimulation
Samples were collected in RNA Lysis buffer from Monarch® Total RNA Miniprep Kit according to the manufacturer's guidelines. RNA Extraction conducted using Monarch® Total RNA Miniprep Kit according to manufactures protocols Human mRNA Sequencing (WOBI) conducted by Novogene. mRNA library preparation (poly A enrichment). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.