GSM8078045: Panc1 shRBM42 Poly RNA 7-10 bio rep 2; Homo sapiens; RNA-Seq
For each sample, equal volumes per polysome fraction were pooled as follows fractions 3-6 for untranslated, fractions 7-10 for low translation and fractions 11-14 for high translation. The RNA from each pool was purified using Trizol-chloroform followed by isopropanol precipitation. Then 1 or 2μl of 1:200 ERCC RNA Spike-In Mix (Invitrogen, 4456740) was added to each fraction pool, depending upon the RNA amount. DNase treatment was performed with TURBO DNase (Ambion, AM2238) for 30 min at 37°C and samples underwent acid phenol extraction and were resuspended in 15μl of Ultra Pure DNase/RNase-Free Distilled Water. 10ng was used as input into the SMARTer Stranded Total RNA-Seq Kit v3-Pico (Takara)