GSM3897536: WT_NPC_1; Homo sapiens; RNA-Seq
For RNA-Seq, total RNA was extracted by TRIzol and subjected to quality control and quantification; For ChIP-Seq, cross-linked cells were lysed in lysis buffer, and the samples were sheared for 14 cycles (30s on/off) in a Bioruptor Pico (Diagenode, Belgium) to achieve an average fragment size of 200-300 bp. Solubilized fragmented chromatin was immunoprecipitated with Anti-FLAG® M2 Magnetic Beads (M8823, Sigma-Aldrich). Antibody-chromatin complexes were pulled down using a magnetic separation rack, washed several times, and then eluted from the magnetic beads. Reverse crosslink was performed subsequently under 65℃ for at least 4 hours. ChIP-DNA was treated with RNase A and Proteinase K, precipitated with ethanol. ChIP-DNA was finally solved in nuclease-free water and quantified using Qubit (Thermo). For RNA-Seq, cDNA libraries were generated using the NEBNext Ultra DNA Library Prep Kit for Illumina (E7370S, NEB). High-throughput sequencing was performed on a HiSeq2500 instrument at the Berry Genomics Co, Ltd, using a 125 bp paired-end-reads setting; For ChIP-Seq, sequencing libraries were generated by using the NEBNext Ultra DNA library preparation kit (E7370, NEB). Libraries were quality-controlled and quantified using a Qubit 2.0 Fluorometer (Life Technologies) and Agilent 2100 Bioanalyzer (Agilent Technologies). High-throughput sequencing was performed on a Illumina HiSeq instrument at the Berry Genomics Co, Ltd.
GEO
GSM3897536