GSM2965770: iDF_mH2A2GFP_0h_RNA-seq_rep3; Mus musculus; RNA-Seq
Cells were grown to less than 80% confluency in 6-well dishes for RNA isolation. Total RNA was extracted using RNeasy Mini Kit (Qiagen) according to manufacturer's protocol. Quality control of extracted RNA was performed using Agilent RNA 6000 Nano Kit and all samples have RNA Integrity Number (RIN) higher than 9.8. Poly(A) mRNA was selected from 1.5 ug total input RNA using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB) according to manufacturer's protocol. Directional, strand specific RNA libraries were prepared using the NEXTflex Rapid Directional RNA-seq Kit (Bioo Scientific) according manufacturer's protocol. The quality of the library was analyzed by Agilent bioanalyzer. The barcoded libraries were subject to 80-bp, single-end sequencing with an Illumina NextSeq 500 instrument.