GSM5904800: siGATA3#2; Homo sapiens; RNA-Seq
Total RNA was isolated from cells using RNeasy Mini kit (QIAGEN, #74104) and was treated with DNase I (QIAGEN #79254). RPFs without ribosomal RNA were run on a 15% Urea denaturing-PAGE gel, and the gel slides from 28nt~30nt were excised. The RPF RNAs were eluted and precipitated followed. The genomic DNAs were extracted by using QIAamp DNA Mini Kit (QIAGEN). RNA-seq libraries were prepared from 2 μg of total RNA, using TruSeq Stranded mRNA Library Prep kit (Illumina # 20020594), according to the manufacturer's instructions.  Sample preparation for ribosome profiling was conducted according to the manufacturer's specifications for the TruSeq Ribo Profile (Mammalian) Library Prep Kit (Illumina).