Author: Ian M. Silverman; Sager J. Gosai; Nicholas Vrettos; Shawn W. Foley; Nathan D. Berkowitz; Zissimos Mourelatos; Brian D. Gregory
Title: Isolation and sequencing of AGO-bound RNAs reveals characteristics of mammalian stem-loop processing in vivo Document date: 2018_4_6
ID: 1pbshnw9_27
Snippet: mRNA-seq was performed as previously described (37) . Briefly, total RNA was purified from the HEK293T cell cultures (miRNeasy; Qiagen). Poly(A) + RNA was isolated using oligo(dT) beads (Life Technologies). RNA was fragmented for 7 minutes using Fragmentation Reagent (Life Technologies). mRNA-seq libraries were then generated using the Illumina mRNA-seq kit (Illumina). Reads were trimmed with Cutadapt, mapped with Tophat2, and gene expression was.....
Document: mRNA-seq was performed as previously described (37) . Briefly, total RNA was purified from the HEK293T cell cultures (miRNeasy; Qiagen). Poly(A) + RNA was isolated using oligo(dT) beads (Life Technologies). RNA was fragmented for 7 minutes using Fragmentation Reagent (Life Technologies). mRNA-seq libraries were then generated using the Illumina mRNA-seq kit (Illumina). Reads were trimmed with Cutadapt, mapped with Tophat2, and gene expression was quantified using HTseq (38) (39) (40) . DEseq2 was used to perform differential expression analysis (41)
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