GSM1891968: RPF-siKrr1-2; Mus musculus; OTHER
RNA-seq and Ribosome profiling libraries were prepared as described previously (Cho et al. 2012), according to the protocol of ARTseq Ribosome Profiling Kit (Epicentre) with minor modifications. Briefly, cell lysates were divided equally for RNA-seq library and ribosome profiling library. Lysates were digested by RNase I (Ambion) and gel purified in the range of 30 nt for ribosome profiling or 40-60 nt for RNA-seq library. The purified RNAs were subjected to reverse-transcription, after ligated with 3′ and 5′ adapter. The cDNA was used as a template for PCR amplification using Phusion polymerase (NEB). Reactions were separated by acrylamide gel and purified again. Quantify and characterize the purified cDNA libraries using bioanalyzer (Agilent) and Kapa library quantification kit, respectively. The libraries were sequenced on Illumina HiSeq 2000 (multiplexing 50 base single-end). All adapters and primers were synthesized by IDT.
GEO Accession
GSM1891968