title: GSM3371781: rna_seq_wt_minus_r2; Mus musculus; RNA-Seq
library_construction_protocol: For Ribo-seq, sorted erythroblasts were washed twice with cold phosphate-buffered saline and then treated with cycloheximde (100 ug/ml) at 37℃ for 5 min, followed by isolation of ribosome protected fragments (RPFs) using ARTseq-Ribosome Profiling Kit (Illumina). For mRNA-seq, total RNAs were extracted by RNeasy Plus kit (Qiagen) and polyA RNAs were isolated using an Oligotex mRNA kit (Qiagen). Ribo-seq cDNA libraries were prepared as described (Ingolia et al. 2009, Ingolia, Lareau, and Weissman 2011). Briefly, RPFs were dephosphorylated and linkers were ligated using T4 RNA ligase. RNA samples were then reverse transcribed, circularized and PCR amplified for 12 cycles. PCR products were subjected to gel purification before sequencing. mRNA-seq cDNA libraries were prepared by the MIT BioMicro Center.
attributes.tag: GEO Accession
attributes.value: GSM3371781