GSM9244853: MK801 device, pooled fraction, rep 3; Homo sapiens; RNA-Seq
For control devices, invasive cells were physically separated from the non-invasive “core” cells using a scalpel. For MK-801 treated devices, the invasive fractions were not large enough to isolate separately from the core, so both fractions were pooled for these samples.  Cells were isolated by degrading the hydrogel with hyaluronidase and centrifuging to get a cell pellet. RNA was then extracted using Trizol (Invitrogen) according to manufacturer's instructions. Isolated RNA was sent to Novogene for library construction, quality control, and sequencing. In brief, mRNA 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 a Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection.