title: RNA-seq of ETCs from barley grains
design: Barley grains from wild type plants and RNAi-lines L4_9 and 16_6 were harvested at 4/5 and 7 DAF, immediately frozen in liquid nitrogen and transferred to a cryostat (-20C). Middle parts of grains were cut by a razor blade, glued onto sample plates by O.C.T medium and serial cross sections (20 m) were mounted on RNAse-free PEN membrane slides (MMI). PEN membrane slides were stored for 7 days at -20C until complete dryness. The MMI Laser Cell Cut system was used to isolate the central endosperm transfer cell (ETC) region. Usually between 60 and 80 ETC regions from 5 different grains were pooled for each biological replicate. Three biological replicates were generated from each genotype at two time points (total number of samples = 18).