GSM7846270: U87,circPLOD2b-NC, rep2; Homo sapiens; RNA-Seq
Human GBM cell U87knocking down of circPLOD2a and circPLOD2b were planted in 6-cm dish respectively and treated in normoxia  (21% O2, 74% N2 and 5% CO2) . 48 hours later, the DMEM medium was removed, and 1mL PBS buffer was added into each dish to wash medium residue. Remove all the PBS buffer from the dishes and add 1mL Trizol buffer. Solution from each dish was collected into 1.5mL EP tube, and stored in liguid nitrogon for further analysis. RNA purity was checked using the NanoPhotometer® spectrophotometer (IMPLEN, CA, USA). RNA concentration was measured using Qubit® RNA Assay Kit in Qubit® 2.0 Fluorometer (Life Technologies, CA, USA). And, RNA integrity was assessed using the RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA). Sequencing libraries were generated using NEBNext® Multiplex Small RNA Library Prep Set for Illumina® (NEB, USA.) following the manufacturer's recommendations. The first-strand cDNA was synthesized using M-MuLV Reverse Transcriptase (RNase H– ). PCR amplification was performed using LongAmp Taq 2X Master Mix, SR Primer for Illumina, and index (X) primer. PCR products were purified and harvested.