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For most RNA FISH experiments, the long-intron PCR products (PCR primer sequences are provided in S1 Table) were cloned into pCRII-TOPO vector (Invitrogen); prepared plasmids were linearized prior to in vitro transcription using SP6 or T7 polymerases (Roche) for production of sense- or antisense-specific probes with incorporation of DIG-16-dUTP or Biotin-16-dUTP (Sigma).
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