RNA-Seq of trabecular meshwork cells: Sciencell-GSK 2
HTM cells (p5) from Sciencell were divided into two groups. The TRPV4-specific agonist group was treated with GSK1016790A (25nM) for 12 hours, while the non-agonist parallel control cultures were incubated under the same conditions without the addition of GSK.After sample quality control, mRNA is isolated using Oligo(dT) Beads. The mRNA is fragmented, and the first cDNA strand is synthesized using the First Strand Synthesis kit. The second strand is synthesized with the Second Strand Synthesis Mix. After end repair and adding an "A" base, RNA-Seq adapters are ligated. The ligated products are purified and amplified by PCR, followed by magnetic bead purification to recover 300 bp-600 bp fragments for sequencing.