AD2935_AW8507_sh_NT4_p
Each sample was collected by scrapping the cells from a 10cm cell culture dish. Pellet was stored at -20 degrees. AW13516, AW8507 and CAL27 cells were cultured in DMEM medium. The media was supplemented with 10% FBS, 2.5 mg/ml Amphotericin-B (Abbott) and 1.25 l/ml Gentamycin (Abbott). The cells were cultured at 37 C in a 5% CO2 incubator. RNA was extracted using the TRIzol solubilization and extraction protocol, as recommended by the manufacturer. Transcriptome sequencing libraries were constructed by using the TruSeq RNA library protocol (Illumina). Briefly, mRNA was purified from 4 g of intact total RNA using oligodT beads and library preparation was done as per the manufacturer's instructions (TruSeq RNA Sample Preparation Kit, Illumina).