characteristics.text: SAMN00153097
characteristics.characteristic: External Id
characteristics.text: LIBEST_021376
characteristics.characteristic: External Id
characteristics.text: University of California
characteristics.characteristic: INSDC center name
characteristics.text: 2007-07-03T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:45:16.863Z
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: IT84S-2049
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005241
characteristics.characteristic: cultivar
characteristics.text: Seeds of IT84S-2049 were from a single plant with a long history of single seed descent (Ehlers). The types of tissue were: 1) germinated seed hypocotyl and primary root, 2) germinated seed epicotyl and foliage leaves, 3 & 4) young trifoliate leaves and apical meristem (one well watered, one drought stressed). For germinated seed samples (1 & 2), seeds were surface sterilized 5 minutes in 1% sodium hypochlorite with 0.01% Tween 20, rinsed with water, germinated on sterile filter paper with water containing 100 units/ml nystatin and 50 microgram/ml cefotaxime, incubated 4-5 days in the dark at room temperature (Fenton). For samples 3 & 4, seeds were planted in 3 gallon pots and kept watered in a glasshouse until plants were ~25 cm in height (~3 weeks)(Fenton). For drought stress (sample 4), water was withheld until visible signs of drought stress occurred (reduced growth relative to watered plants, wilted leaves, less green color), generally ~3 weeks. Total RNA was purified using a TRIzol method (Fenton). RNA was pooled to establish one highly complex RNA sample with a total of 1000 microgram, as follows: 333 microgram of hypocotyl & primary root (sample 1), 333 microgram of epicotyl and foliage leaves (sample 2) and 167 microgram each of stressed and unstressed young trifoliate leaves and apical meristem (samples 3 & 4)(Fenton). PolyA RNA was purified using Promega PolyATtrack mRNA Isolation System IV (Fenton). Five microgram of poly(A) mRNA was used for cDNA synthesis. A primary cDNA library was produced using a Stratagene lambda ZAP XR cDNA Synthesis Kit (Fenton). 250 thousand pfu from the primary library were mass excised to produce a phagemid population (Fenton). Phagemids were plated (Fenton, Resnik), plasmid DNA purified, cDNA clones archived, and DNA sequences determined bi-directionally using an ABI3730 at the Genomics Core Instrumentation facility, University of California, Riverside (Choi). Trace files were downloaded (Close) and then processed (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 with annotations (Close).
characteristics.characteristic: description
characteristics.text: Seedling
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PO_0007131
characteristics.characteristic: dev-stage
characteristics.text: E. coli TJC121
characteristics.characteristic: lab-host
characteristics.text: Vigna unguiculata
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_3917
characteristics.characteristic: organism
characteristics.text: EcoRI
characteristics.characteristic: re_1
characteristics.text: XhoI
characteristics.characteristic: re_2
characteristics.text: mixed
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001271
characteristics.characteristic: tissue-type
characteristics.text: Cowpea IT84S-2049 cDNA Library UCRVU03-1
characteristics.characteristic: title
characteristics.text: Phagemid pBluescript SK(-)
characteristics.characteristic: v_type
characteristics.text: Lambda Uni-ZAP XR, excised phagemid
characteristics.characteristic: vector