Illumina HiSeq 2500 sequencing; GSM1638578: rea1 mutant total; Zea mays; RNA-Seq

rea1 mutant total

Analysis of  gene expression level. The hypothesis tested in the present study was that rea1 mutant influence the translation level of translation and nucleosome assembly associated genes. Results provide important information of the gene translation level regulation of ribosome proteins, elongation factors, histones and genes associated with other biological processes. Overall design: Endosperm total and polysome-bound mRNA profiles of 15DAP wild type (WT) and rea1 mutant were generated by deep sequencing, in triplicate, using Illumina Hiseq 2500 (Zea mays).

Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and rea1 mutant total and polysome-bound Transcriptomes

Submitted by Gene Expression Omnibus on 25-APR-2015

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