characteristics.text: SAMN00156414
characteristics.characteristic: External Id
characteristics.text: LIBEST_001849
characteristics.characteristic: External Id
characteristics.text: Washington University School of Medicine
characteristics.characteristic: INSDC center name
characteristics.text: 1999-06-14T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:35:52.603Z
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: Williams
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0007023
characteristics.characteristic: cultivar
characteristics.text: The mRNA was isolated from entire roots of 2-month-old 'Williams' plants that were greenhouse grown in 5-gallon pots. To suppress nodulation, Black Gold All-Purpose potting soil was supplemented with: 0.36g/L available phosphoric acid (P2O5), 20mg/L urea N, 0.16g/L S, 0.49mg/L B, 2.5mg/L Cu, 0.15g/L Fe, 13.53mg/L Mn, 0.26mg/L Mo, 14mg/L Zn, 20mg/L Ca, and the following nutrients in a slow-release form (Osmocote): 0.165g/L ammonia N, 0.185g/L nitrate N, 0.35g/L available phosphoric acid, and 0.35g/L soluble potash. No nodules were visible on the roots at harvest. Stratagene's cDNA Synthesis Kit (catalog #200401) was used to synthesize the cDNA. First-strand synthesis was performed with 5-methyl dCTP, hence the ligated cDNA is hemimethylated. Stratagene's first-strand synthesis primer was used [GAGAGAGAGAGAGAGAGAGAACTAGTCTCGAG(T)-18]. After second-strand synthes is, the cDNA ends were 'polished' with clone Pfu DNA polymerase, ligated to EcoRI adapters, and phosphorylated. The XhoI site within the first-strand synthesis primer was restricted by digestion with XhoI; all XhoI sites in the cDNA would be protected by their hemimethylated status. The cDNA constructs were size-fractionated with a 400bp cutoff, using a SizeSep 400 Spun column from Pharmacia. The column eluent was then ligated into Stratagene's pBluescript II XR Predigested vector (pBluescript II SK(+) that had been digested with EcoRI and XhoI, and phosphorylated). Both the white and blue colonies appear to contain recombinant plasmids with cDNA inserts. This library was constructed by Dr. Paul Keim and Dr. Virginia Coryell.
characteristics.characteristic: description
characteristics.text: XL10-Gold
characteristics.characteristic: lab-host
characteristics.text: Glycine max
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_3847
characteristics.characteristic: organism
characteristics.text: EcoRI
characteristics.characteristic: re_1
characteristics.text: XhoI
characteristics.characteristic: re_2
characteristics.text: Gm-c1009
characteristics.characteristic: title
characteristics.text: plasmid
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/SO_0000155
characteristics.characteristic: v_type
characteristics.text: pBluescript II XR
characteristics.characteristic: vector