GSM8646252: Donor1, sgLAYN; Homo sapiens; RNA-Seq
Treg cells were isolated respectively by gating on live CD45+CD4+CD25+LAYN+ or CD45+CD4+CD25+LAYN+ cells. Cells were sorted into fresh media, plate and keep in culture for 24h. After 24h cells were pellet then flash frozen on dry ice before being processed by Novogen. 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 prepared at and by Novogen. Library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. Then checked with Qubit and real-time PCR