RNA-seq of cotyledons: 7-short-day old Arabidopsis seedling
Cotyledons and hypocotyls were dissected from 6-day-old seedlings at ZT-1~ZT-1.5 and flash freezing in liquid nitrogen. RNA was extracted  with an RNeasy plant Mini Kit (QIAGEN) with an on-column DNase (QIAGEN) treatment. RNA quality were assessed by capillary electrophoresis (NanoChip, Agilent Bioanalyser). Poly-A RNA was enriched from 500 ng total RNA by the Poly(A) mRNA Magnetic Isolation Module (New England Biolabs). RNA-seq libraries were prepared as described in Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs). A total of thirteen cycles were applied to enrich library concentration. Sequencing-by-synthesis was done on HiSeq 3000 device at the Max Planck Genome-centre Cologne in paired-end read mode 2 x 150 bp.
SUB9946344
4163_E