GSM1477471: large polysome footprint; Drosophila melanogaster; RNA-Seq
We purified mRNAs in small polysomes, away from monosomes (80S), ribosomal subunits (40S, 60S) and large polysomes. 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
GEO Accession
GSM1477471