Nucleated erythrocytes can play more than one tune
mRNA was fragmented using RNase III and fragmentation was verified with an Agilent Bioanalyzer 2100 using the RNA 6000 Pico Chip Kit.  Samples were hybridised (Adaptor Mix A), ligated, and reverse transcribed.  cDNA was subjected to electrophoresis on a Novex 6% TBE-Urea Gel (Invitrogen, USA), stained with SYBR Gold (Invitrogen, USA), and the region corresponding to 100-200 nucleotides was excised from the gel.  In-gel PCR was performed and reactions purified with the PureLink PCR Micro Kit (Invitrogen, USA) according to the manufacturer's protocol and analysed with an Agilent Bioanalyzer 2100 using the DNA 1000 Chip Kit (Agilent).  Sequencing was perform using the SOLiD3 System (Applied Biosystems) according to the manufacturer's protocol. A total of four panels were sequenced corresponding to two panes for each library (stimulated with poly I:C and nonstimulated).