title: Illumina Genome Analyzer IIx sequencing; Tissue layers from developing wheat grain at 12 days post-anthesis
design: Tissue layers from developing wheat grain at 12 days post-anthesis
library_construction_protocol: Immature grain at 12 days post-anthesis were collected from floret positions 1 & 2 of spikelets in the  central 3/4 of the ears. Three tissue layers were isolated by manual dissection: the 'outer pericarp', which included the cuticle, outer epidermis, hypodermis and parenchyma cells; the 'inner pericarp' which comprised the cross cells, inner epidermis, integuments and aleurone; and the endosperm. Tissues were frozen in liquid nitrogen and stored at -80oC Plants growth in 15cm pots of a peat and loam-based compost in a glasshouse: 16 hr day (), 8 hr night (). Plant were tagged at anthesis. RNA was extracted according to Wan et al. (2009) Plant Biotechnol J 7, 401-410. Libraries were prepared using the Illumina TruSeq mRNA sample kit using 4ug of total RNA as described in the  Illumina TruSeq RNA sample preparation guide, Part 15008136 Rev A, Nov 2010.
attributes.tag: Experimental Factor: organism part
attributes.value: outer pericarp