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1st strand cDNA was primed with a Not I - oligo(dT) primer [5' TGTTACCAATCTGAAGTGGGAGCGGCCGCGCTTTTTTTTTTTTTTTTTTT 3'], double-stranded cDNA was size selected, ligated to Eco RI adapters (Pharmacia), digested with Not I and cloned into the Not I and Eco RI sites of a modified pT7T3 vector (Pharmacia). Library went through one round of normalization to a Cot = 53. Library constructed by Bento Soares and M.Fatima Bonaldo. The adult brain RNA was provided by Dr. Donald H. Gilden. Tissue was acquired 17-18 hours after death which occurred in consequence of a ruptured aortic aneurysm. RNA was prepared from a pool of tissues representing the following areas of the brain: frontal, parietal, temporal and occipital cortex from the left and right hemispheres, subcortical white matter, basal ganglia, thalamus, cerebellum, midbrain, pons and medulla.
Nucleotide
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