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Gm-c1004

Identifiers
BioSample: SAMN00156191; EST: LIBEST_001617
Organism
Glycine max (soybean)
cellular organisms; Eukaryota; Viridiplantae; Streptophyta; Streptophytina; Embryophyta; Tracheophyta; Euphyllophyta; Spermatophyta; Magnoliopsida; Mesangiospermae; eudicotyledons; Gunneridae; Pentapetalae; rosids; fabids; Fabales; Fabaceae; Papilionoideae; 50 kb inversion clade; NPAAA clade; indigoferoid/millettioid clade; Phaseoleae; Glycine; Glycine subgen. Soja
Attributes
cultivarWilliams
tissueroot
lab hostXL10-Gold
vectorpBluescript II XR
v_typeplasmid
re_1EcoRI
re_2XhoI
Description

Root cDNA. The mRNA was isolated from entire roots of 8 day old 'Williams' seedlings which were propagated on paper towels with distilled water. 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 synthesis, 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 500bp cutoff, using GibcoBRL Life Technologies' cDNA Size Fractionation column. 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. Blue colonies 9n=15) have been sequenced, and possess putative cDNA inserts. This library was constructed by Dr. Paul Keim & Virginia H. Coryell, Department of Biology, Box5640, Northern Arizona University, Flagstaff, AZ 86011, Phone: 520-523-1078 (Dr. Paul Keim), 520-523-1372 (Virginia H. Coryell), Fax: 520-523-7500, email: paul.keim@nau.edu, virginia.coryell@nau.edu

Submission
Washington University School of Medicine, Shoemaker R/Public Soybean EST Project; 1999-02-01
Accession:
SAMN00156191
ID:
156191

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