GSM3074852: Met-_CHX; Homo sapiens; OTHER
Equal aliquots of the ribosome fractions separated by sucrose gradient were pooled and treated with E. coli RNase I (Ambion) to digest regions of mRNAs not protected by ribosomes. RNAs were extracted using TRIzol LS. cDNA libraries were constructed from the ribosome-protected mRNA fragments by poly(A)tailing and reverse transcription using barcode containing oligonucleotides (one for each treatment condition). For deep sequencing, the cDNA library was amplified by PCR using the Phusion High-Fidelity polymerase and primers containing Illumina cluster generation sequences. Equal amounts of barcoded samples for the three conditions were mixed together, and approximately 3–5 pM of the pooled DNA sample was used for cluster generation followed by deep sequencing.
GEO Accession
GSM3074852