GSM5667165: tracer tRNA replicate 2; Homo sapiens; ncRNA-Seq
Cells were collected from culture dish and solubilized in Trizol. Total RNA was extracted, and separated using UREA-PAGE. tRNAs were excised, eluted from crushed gel pieces and precipitated by isopropanol. Refolded tRNAs were further treated with recombinant angiogenin endonuclease, radiolabeled using P32-gamma-ATP and used to monitor DDX3X helicase unwinding activity Libraries were prepared according to NEBNext small RNA cloning protocol (E7580S)