alias: GSM2210969
title: GSM2210969: Stage_14_wt_No_chx_RPF_7; Drosophila melanogaster; OTHER
library_construction_protocol: Samples were transferred to RPF lysis buffer (10 mM Tris-HCl, pH 7.4, 5 mM MgCl2, 100 mM KCl, 2 mM DTT, 100 µg ml–1 cycloheximide, 1% Triton X-100, 500 U ml–1 RNasin Plus, and protease inhibitor (1x complete, EDTA-free, Roche)), centrifuged at 10000 rpm for 5 min, with obtained supernatants flash frozen in liquid nitrogen to be stored at –80 °C. Thawed lysates were then used for ribosome footprint profiling, or total RNA was extracted with TRIreagent and used for RNA-seq and PAL-seq. Samples Stage_14_wt_No_chx_RNA_1, Stage_14_wt_No_chx_RPF_7, Stage_14_wt_No_chx_PAL_46, Activated_egg_wt_No_chx_RNA_2, Activated_egg_wt_No_chx_RPF_8, and Activated_egg_wt_No_chx_PAL_47 were prepared using lysates made with RPF lysis buffer that did not contain cycloheximidie. Samples were split and prepared for ribosome-footprint profiling and RNA-seq as described (Kronja et al., 2014b), as well as for PAL-seq as described (Subtelny et al., 2014).  Detailed protocols are available at http://bartellab.wi.mit.edu/protocols.html. For each stage, RNA, RPF, and tail length measurements were from the same sample.
identifiers.namespace: GEO
identifiers.id: GSM2210969