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ERX2090098: Illumina HiSeq 3000 paired end sequencing; RNA-seq of enriched barley stomatal complexes versus total leaves
1 ILLUMINA (Illumina HiSeq 3000) run: 45.8M spots, 13.1G bases, 5Gb downloads

Design: RNA-seq of enriched barley stomatal complexes versus total leaves
Submitted by: PGSB - Plant Genome and Systems Biology Helmholtz Center Munich German Research Center for Environmental Health (GmbH) (PGSB - Plant Genome and Systems Biology Helmholtz )
Study: RNA-seq of enriched barley stomatal complexes versus total leaves
show Abstracthide Abstract
We intend to identify transporters and their regulators involved in barley stomatal movement as well as components of the ABA signaling pathway. We isolated epidermal peels where only stomatal guard cells and their subsidiary cells survived, while other epidermal cell were mechanically disrupted and sequenced the resulting RNAs. Thus we analyzed transcripts differentially expressed between the barley stomatal complex and total leaves. These data served as the first overview of genes expressed in the stomatal complex and for cloning of relevant transporters.
Sample: Sample 1
SAMEA104158944 • ERS1817962 • All experiments • All runs
Name: Sample 1_p
Instrument: Illumina HiSeq 3000
Strategy: RNA-Seq
Selection: RANDOM
Layout: PAIRED
Construction protocol: Barley (Hordeum Vulgare cv. Barke) seeds were provided by a commercial supplier (Saatzucht J. Breun GmbH & Co. KG) and cultivated at 22/16 °C and 50 ± 5% RH at a 12/12h day/night cycle and a photon flux density of 400 μmol m-2 s-1 (µE) white light (Philips Master T Green Powers, 400 W). The preparation of epidermal peels was performed using a method developed previously (Shen et al., 2015). Epidermal Peels were collected from the abaxial side of 8 to 10 day old leaves. RNA was extracted from a total of 20 epidermal peels per sample using the NucleoSpin® RNA Plant Kit (Macherey-Nagel, Germany). RNA isolation from whole leaves was performed equally. The extracted RNA was treated with RNase-free DNase (New England Biolabs, Ipswich, MA, USA). Quality control measurements were performed on a 2100 Bioanalyzer (Agilent, Santa Clara, CA, USA) and the concentration was determined using a Nanodrop ND-1000 spectrophotometer (Thermo Fisher Scientific, Wilmington, DE, USA). Libraries were prepared with the TruSeq RNA Sample Prep Kit v2 (Illumina, San Diego, CA, USA) using 1 µg of RNA
Spot descriptor:
forward101  reverse

Experiment attributes:
Experimental Factor: organism part: stomatal complex
Runs: 1 run, 45.8M spots, 13.1G bases, 5Gb
Run# of Spots# of BasesSizePublished


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