GSM8155465: miP-PSTPIP2-solvent_1; Homo sapiens; RNA-Seq
Cells were lysed in a lysis buffer containing Tris-HCl pH7.5 (50mM, Applichem, Germany), NaCl (150mM, Merck, Darmstadt, Germany), MgCl2,(10mM, Invitrogen, Schwerte, Germany), NP-40 (1%, Merck) dissolved in UltraPure DNase/RNase-free DW (Invitrogen) and supplemented with NaPPi (10mM, Merck), NaF (20mM, Applichem, Germany), okadaic acid (10nM, LC laboratory, USA), Na3VO4 (2mM, Sigma-Aldrich), protease inhibitor mix, (12µl/ml, Sigma-Aldrich), PMSF (4µl/ml, Carl Roth, Germany), SUPERase In RNase Inhibitor (200U/ml, Invitrogen), TURBO DNaseI (25U/ml, Invitrogen). Total RNA was purified using the miRNeasy Micro kit (QIAGEN), according to the manufacturer's instructions. RNA and library preparation integrity were verified with LabChip Gx Touch 24 (Perkin Elmer). 0.5-1µg of total RNA was used as input for Corall Total RNA-Seq Library Prep Kit following manufacture's instructions (Lexogen). Sequencing was performed on the NextSeq2000 instrument (Illumina) using P3 flowcell and 1x72bp single end setup.