GSM3267694: SWngfQT4; Mus musculus; RNA-Seq
Ribosome bound RNA fragments and corresponding sets for total RNA were isolated as described in Ingolia et al., 2012 and Hornstein et al., 2016 with minor adjustments in the composition of the polysome lysis buffer (20 mM Tris-HCl, pH 7.5, 250 mM NaCl, 15 mM MgCl2, 1 mM DTT, 0.5% (vol/vol) Triton X-100, 2.5 U/ml DNase I, 40U/mL RNasin and 50ug/mL emetine). MicroSpin S-400 columns (GE Healthcare) were used to isolate ribosome bound RNAs. After rRNA was removed using RiboCop rRNA depletion kit (Lexogen), footprints were dephosphorylated then size selected (28-34nt) by PAGE on 15% TBE-Urea gels (Bio-Rad). Footprints and total RNA libraries were generated by SMARTer smRNA-Seq kit for Illumina (Clonetech) and Quantseq 3ʹ mRNA-Seq library kit (Lexogen) respectively.
GEO Accession
GSM3267694