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Zebrafish embryos were injected with 4-thiouridine (4sU; 1.5 mM) at the 1-cell stage and development proceeded for a defined time (Δt), followed by total RNA preparation and SLAMseq employing 3′ mRNA sequencing. Zygotic transcripts were distinguished from maternal mRNAs by the presence of T>C conversions at sites of 4sU incorporation as identified by SLAMDUNK.21

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