characteristics.text: SAMN00166067
characteristics.characteristic: External Id
characteristics.text: LIBEST_013887
characteristics.characteristic: External Id
characteristics.text: University of California
characteristics.characteristic: INSDC center name
characteristics.text: 2003-06-20T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:41:40.290Z
characteristics.characteristic: INSDC last update
characteristics.text: live
characteristics.characteristic: INSDC status
characteristics.text: Generic
characteristics.characteristic: NCBI submission model
characteristics.text: Generic.1.0
characteristics.characteristic: NCBI submission package
characteristics.text: Dicktoo
characteristics.characteristic: cultivar
characteristics.text: Seeds of barley (Hordeum vulgare L. cv. Dicktoo) were germinated in dishpans containing UCR-mix soil. Seedlings were kept in a growth chamber at 20C (day/night) and allowed to grow at 60-70% soil moisture content. After 3-4 days, the water was withheld in order to apply drought until the soil moisture content was reduced to 10-12%, which took another 3-4 days. At the time of extraction of epidermis, leaf water and osmotic potentials and extracted lower epidermis osmotic potentials had dropped to 15.9%, 20.2% and 24.7% of controls, respectively, as determined using a vapor pressure osmometer (Model 5100C, Wescor, Inc., Logan, UT). Epidermal strips were quickly peeled off of seedlings and immediately submerged in liquid nitrogen. About 15-20 g of epidermal tissue was collected and used to extract total RNA. Total RNA was extracted using Concert Plant RNA Reagent (Invitrogen 12322-012). Poly(A) RNA was purified using PolyATrack mRNA Isolation System IV (Promega). A cDNA library was made using a Uni-ZAP cDNA synthesis kit (Stratagene). A total of 1 million primary lambda cDNA clones were mass-excised in vivo to give a population of pBluescript SK(-) phagemids. All steps to this point were performed in the TJ Close lab at the University of California, Riverside by A. Wahid with some assistance from R.D. Fenton. Phagemids were plated on the TJC121 host strain, plasmid DNA purified, cDNA clones archived, and DNA sequences determined using the T7 primer (mainly 3' end reads) using an ABI3730 at the Arizona Genomics Institute, University of Arizona (Collura, Feuerbacher, Kim, Kudrna, Wing, Yu). Chromatogram files were transmitted to UC Riverside (by Yu), then processed at UC Riverside (by Wanamaker) using the HarvEST pipeline (http://harvest.ucr.edu) to remove vector and cloning oligo sequences and various contaminants, and to trim to a high quality region. Sequences that retained a phred 17 region of at least 100 bases were deposited to GenBank.
characteristics.characteristic: description
characteristics.text: 1-2 week seedlings
characteristics.characteristic: dev-stage
characteristics.text: E. coli TJC121
characteristics.characteristic: lab-host
characteristics.text: Hordeum vulgare subsp. vulgare
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_112509
characteristics.characteristic: organism
characteristics.text: EcoRI
characteristics.characteristic: re_1
characteristics.text: XhoI
characteristics.characteristic: re_2
characteristics.text: lower leaf epidermis
characteristics.characteristic: tissue-type
characteristics.text: Drought-stressed Dicktoo barley epidermis cDNA library
characteristics.characteristic: title
characteristics.text: Phagemid pBluescript SK(-)
characteristics.characteristic: v_type
characteristics.text: Lambda Uni-ZAP XR, excised phagemid
characteristics.characteristic: vector