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10.1096/fj.14-259598 20
Physiological and pathophysiological roles of excitatory amino acids during central nervous system development
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In vitro transcription to prepare biotinylated PRKAA1 5UTR and scrambled RNA probes PRKAA1 5UTR and scrambled control RNA probes were synthesized using BamHI-linearized pUC57-T7 templates in an 80-l reaction buffer containing 5 transcription buffer (Roche, RPOLT7-RO), 8 l of 10 Dig labeling mixture (Merck, 11277073910), 8 l of 0.1 M DTT (Sigma, D9163