GSM3957961: MB002_GFP_3; Homo sapiens; RNA-Seq
The total RNA is then extracted from the remaining lysate using Trizol LS (Invitrogen). The polysome fractions are collected and pooled for polyribosome associated RNA extraction using Trizol LS. Extracted polysome associated RNA is resuspended in 50μL of sterile ultrapure water. 1ug of total RNA and an equivalent volume of RNA from the polysome fraction were used for library preparation as per Illumina standard RNA-seq protocol. An equal amount of Spike ins RNA Variant Controls (SIRVs, Lexogen SIRV-Set3 and ERCC) were added to each sample prior to library preparation to normalize for sequencing bias and to determine the threshold for measurable statistics. The library was then used to build clusters on the Illumina flow cell and analysis was done using Illumina Hiseq 4000 platform (Illumina, San Diego, CA, USA) at the UCSD Institute for Genomic Medicine to a target depth of 30M reads per sample.
GEO Accession
GSM3957961