Pltlts.4N4.Long.rRNAdpltd
Highly purified, leukocyte-depleted platelet (LDP) preparations were obtained as previously described (Nagalla et al., 2011). RNA extraction was performed with TRIzol¨ (Invitrogen, Carlsbad, CA) and RNA quality was assessed by the Agilent bio-analyzer PICO chip. In some experiments, 2 ¼g of total RNA was depleted of large 18S and 28S rRNA as well as 5S and 5.8S using the RiboMinus Eukaryotic Kit (Invitrogen), which uses biotinylated probes designed against these rRNAs (following the manufacturer's protocol). Total RNA and rRNA-depleted RNA were fragmented using RNAse III digestion for 13 min in a 10 ¼l reaction containing 1 ¼l of 10X RNAse III buffer and 20 U of RNAse III. After incubation the RNA was purified using the RiboMinus Concentration Module (Invitrogen), and the size and yield of RNA was determined using the Agilent bio-analyzer PICO chip.   Library construction, emulsion PCR, workflow analysis and sequencing runs were performed following standard AB/Life Technologies protocols.