GSM2100591: M phase RNA-seq oligo-dT enriched replicate 2; Homo sapiens; RNA-Seq
Total RNAs were extracted by TRIzol (Invitrogen) For oligo-dT enriched mRNA-seq, total RNA extracted by TRIzol (Life Technologies) was treated with DNaseI (Takara) and purified by RNeasy Minelute Cleanup kit (QIAGEN). mRNAs were enriched by oligo dT capture. 5 μg of RNAs were fragmented by incubating in the fragmentation buffer provided by NEBNext Magnesium RNA Fragmentation Module at 95 °C for 6.5 min. Fragmented RNAs were size-selected for 60-80 nt RNAs, dephosphorylated by Antarctic phosphatase (NEB) and 5′ phosphorylated by PNK reaction (Takara). RNAs were then subjected to the library preparation protocol used in ribosome profiling described above. The cDNA libraries were sequenced by Illumina HiSeq (1 × 51 bp single run).
GEO
GSM2100591