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UI-R-DK0

Identifiers
BioSample: SAMN00164153; EST: LIBEST_009642
Organism
Rattus norvegicus (Norway rat)
cellular organisms; Eukaryota; Opisthokonta; Metazoa; Eumetazoa; Bilateria; Deuterostomia; Chordata; Craniata; Vertebrata; Gnathostomata; Teleostomi; Euteleostomi; Sarcopterygii; Dipnotetrapodomorpha; Tetrapoda; Amniota; Mammalia; Theria; Eutheria; Boreoeutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; Murinae; Rattus
Attributes
strainSprague-Dawley
development stageADULT
lab hostDH10B (Life Technologies)
vectorpT7T3D-PacI
v_typePhagemid
re_1Not I
re_2Eco RI
Description

The UI-R-DK0 library is a subtracted library derived from a mixture of five individually tagged normalized rat libraries: brain-nRBP (20%), heart-nRHP (20%), kidney-nRKP (20%), aorta-nRAP (20%), and placenta-nRPP (20%). Each original library was constructed from a mixture of equal amounts of RNA from seven different developmental time-points: embryonic day 17, embryonic day 19, embryonic day 21, adult day 1, adult day 12, adult day 75, and adult day 200. (Exception: the aorta pool does not contain embryonic day 17 RNA and the placenta pool contains only the three embryonic stages). Each library was normalized individually according to the procedure described by Bonaldo, Lennon & Soares (Genome Research Genome 6: 791-806, 1996). For construction of the DK0 subtracted library, plasmid DNA from each of the five individually tagged normalized libraries was mixed in the proportions specified above and electroporated into competent bacteria for production of single-stranded circular DNA representing the pool of libraries. Single-stranded circular DNA representing these five normalized libraries was then used as a tracer in a subtractive hybridization with a driver (PCR amplified inserts from a plasmid DNA template preparation) comprising: a) a set of about 1,000 arrayed clones from each of the five non-normalized libraries of brain (CT0s), heart (CS0s), kidney (CU0s), aorta (CW0s), and placenta (CX0s). The resulting pool of approximately 5,000 clones represented about 33.3% of the final driver population. A set of about 2,000 arrayed clones from each of the five normalized libraries of brain (CT0), heart (CS0), kidney (CU0), aorta (CW0), and placenta (CX0). The resulting pool of about 10,000 clones represented about 66.6% of the final driver population.

Submission
Children's Memorial Research Center, Soares, MB; 2001-07-19
Accession:
SAMN00164153
ID:
164153

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