GSM4435507: S2_polysome; Drosophila melanogaster; OTHER
We purified mRNAs in polysomes, away from monosomes (80S), ribosomal subunits (40S, 60S). Footprinting was performed overnight at 4°C with RNaseI (Invitrogen), stopped with SUPERase·In TM RNase inhibitor (Invitrogen) and precipitated. mRNA from total cytoplasmic lysate, was purified using oligo (dT) Dynabeads (Invitrogen) and fragmented by alkaline hydrolysis. 28-34 nt ribosome footprints and 50-80 nt mRNA fragments were gel purified and prepared as previously described (Ingolia et al, 2012; Ingolia et al, 2009; Ingolia et al, 2011) for Next Generation Sequencing. Libraries were sequenced on Illumina NextSeq and MiSeq machines with 50, 75 and 150bp SingleEnd read protocol.
GEO Accession
GSM4435507