GSM5952499: Y27-E6D7_5; Homo sapiens; RNA-Seq
oligo-dt primers; Samples were harvested and lysed in TCL lysis buffer (Qiagen). Lysates were transferred to TurboCapture 384 format plates (Qiagen) and incubated at RT 1 hr at 100 rpm for polyA RNA capture. Cell lysate then washed with a binding buffer (20 mM Tris-HCl, pH 7.5 at 25 ⁰C, 500 mM NaCl, 1 mM EDTA, 0.1% SDS) twice and 20 µL probes mixture was added into each well with 0.05 pmol/oligo in binding buffer. The binding mixture was incubated at 65 ⁰C for 10 minutes and gradually the temperature was lowered to 45 ⁰C by every 3 degree/per minute rate before 1 hr incubation at 45 ⁰