GSM4676911: 4T1_Ctrl2; Mus musculus; RNA-Seq
Total RNAs from Ctr, vCas9, FAS54, and FAS69 cells were extracted using TRIzol (Life Technology, 15596018). mRNAs were enriched using Oligo d(T)25 Magnetic Beads (NEB S1419S) following manufacturer's protocol. mRNAs were then fragmented at 94 °C in 10mM MgCl2 buffer. Fragmented RNAs were end-repaired by T4 PNK (NEB M0201S) and Poly(A)-tailed by  E. coli Poly(A) Polymerase (NEB M0276S). The poly(A) tailed RNA fragments were reverse transcribed to cDNA using custom-designed oligo(dT) and locked nucleic acid (LNA) based on SMART-seq2 method (Picelly et al. Nature Protocols, 2014). The cDNAs were then PCR amplified using KAPA HiFi HotStart master mix (KK2601). 200-400 bp fragments of the library were cut from gels for Illumina Hiseq sequencing.
GEO Accession
GSM4676911