GSM1638579: wildtype total; Zea mays; RNA-Seq
For isolation of polysomal RNA, pulverized tissue powder was hydrated in two volumes of polysome extraction buffer (PEB), homogenized, filtered through two layers of sterile Miracloth, and cleared by centrifugation. The supernatant was layered over a 1.75 M sucrose cushion, and centrifuged at 170,000 g, at 4 ℃ for 3 h. The polysome pellet was washed with sterile water and resuspended in 700 μl PEB lacking heparin and detergents. Total or polysome-bound RNA was precipitated from total supernatant or ribosome fraction of same amount of sample powder, by addition of 2.5 vol of 8 M guanidine chloride and 3.5 vol of 99% (v/v) ethanol, and extracted by use of Qiagen Plant RNeasy mini kit. RNA libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM1638579