GSM4971870: Astrocyte WT-1 p53 CHIP; Homo sapiens; ChIP-Seq
For Me-RIPseq, cells samples were lysed in Trizol reagent (Invitrogen, 10296010). mRNA were isolated from total RNA by using mRNA isolation kit (11741985001 Roche). mRNA were fragmented by 10 mM Zncl2 solution and incubated with protein A/G dynabeas and equal amount of m6A antibody (SYSY) to mRNA. Input mRNA were not with antibody incubation. After wash, RNA were extracted by using TrizOL For RNAseq, cells samples were lysed in Trizol reagent (Invitrogen, 10296010).ChIP-seq, cells were fixed, quenched, sonicated to ~250 bp in 1% SDS lysis buffer, and incubated with Protein A/G Dynabeads and 5 µg of the indicated antibody. 2% pre-cleared chromatin prior to primary antibody addition was kept as input DNA. After wash, treatment with RNAse A and Proteinase K, DNA was eluted and purified with Qiagen's PCR purification kit (28106). For RNASeq, library construction was performed using NEBNext Ultra II RNA Library Prep Kit (E7770) following manufacturer's instructions. For ChIPSeq, library construction was performed using KAPA Hyper Prep Kit (KK8500) according to manufacturer's conditions. the poly(A) selection step and starting with the RNA fragmentation step.Libraries were submitted to PE150 high-throughput sequencing.