characteristics.text: SAMN00152347
characteristics.characteristic: External Id
characteristics.text: LIBEST_020558
characteristics.characteristic: External Id
characteristics.text: University of California
characteristics.characteristic: INSDC center name
characteristics.text: 2006-10-30T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:44:52.930Z
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: CB27
characteristics.characteristic: cultivar
characteristics.text: Seeds were germinated in pots (Ehlers) and grown in controlled glass house conditions under natural photoperiod and day/night temperatures of 35/18oC. The growing axillary buds from 10 day old seedlings were harvested from one set of pots, flash frozen and stored at -80oC. Another set of plants was allowed to grow under drought stress induced by withholding water. The growing axillary buds were harvested from 4-day drought stressed seedlings, then flash frozen (Sudhakar). The frozen samples were used for RNA extraction by a TRIzol method (Sudhakar, Mandal, Fenton). A total of 500 microgram each from two pools of three stressed and three unstressed sub-samples were combined to total 1000 microgram of total RNA. mRNA was isolated using PolyATtrack (Promega). Five micrograms of poly(A) mRNA was used for cDNA synthesis (Stratagene). A primary cDNA library was produced using a lambda ZAP XR cDNA Synthesis Kit(Stratagene). One million pfu from the primary library were mass excised to produce a phagemid population (Sudhakar). Phagemids were plated, plasmid DNA purified and sequences studied by students (Finaldi, Pottorff, Pena, Hasegawa, Hasegawa, McCoy, Ramirez, Wong, Alzubi, Ngo) and the teaching assistant (Schroeder) in the Fall 2006 'Plant Genomics and Biotechnology' class taught by Close at UC Riverside. Plasmids were readied for sequencing (Close, Resnik) and DNA sequences determined bi-directionally using an ABI3730 at the Genomics Core Instrumentation facility, University of California, Riverside (Choi). Sequences were 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.
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: growing axillary bud
characteristics.characteristic: tissue-type
characteristics.text: Cowpea CB27 Axillary Bud cDNA Library - UCRVU02-2
characteristics.characteristic: title
characteristics.text: Phagemid pBluescript SK(-)
characteristics.characteristic: v_type
characteristics.text: Lambda Uni-ZAP XR, excised phagemid
characteristics.characteristic: vector