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Protocol publication

[…] RNA was isolated from whole tissue using RNAqueous Micro Kit (Ambion). Total RNA samples were DNase-treated (Ambion), prior to cDNA synthesis using SuperScript reagents (Invitrogen, CA). SYBRgreen qPCR was performed using 1 ng of cDNA and primers designed using Primer3 and Beacon Designer software or found using PrimerBank (http://pga.mgh.harvard.edu/primerbank/). Primer sequences are listed in and . Melt-curves and primer efficiency were determined as previously described (). Gene expression was normalized to the housekeeping gene S29 (Rps29) for mouse and GAPDH for human and to the corresponding experimental control. Reactions were run in triplicate and experiments performed two to three times. […]

Pipeline specifications

Software tools Primer3, Beacon Designer
Databases PrimerBank
Application qPCR
Organisms Homo sapiens, Mus musculus