For quantitative RT-PCRs (Fig. S1G), RNA was extracted from whole embryo lysates at 2 dpf using the Directzol RNA mini kit (Zymo). cDNA was synthesised and amplified using Thermo Fisher SuperScript II Reverse Transcriptase Kit. qPCR was performed on an Applied Biosystems Viia 7 384-well real-time PCR machine. Fold changes were calculated relative to housekeeping genes rpl13 and hprt.
For RT-PCRs during maternal-zygotic transition (Fig. S2A), RNA was extracted from 20 wild-type embryos (pooled) at the one-cell stage, 1 hpf, 3 hpf, 1 dpf and 2 dpf. cDNA was synthesised and amplified using Thermo Fisher SuperScript II Reverse Transcriptase Kit. We included previously verified control genes (Harvey et al., 2013 (link)), including; cyclinb1 (maternally provided), C18H16orf7 (maternally provided, but not transcribed after zygotic transition) and tal1 (not maternally provided). Primers sequences were as follows: tln1 fwd, 5′-GGTCTCATTTCAGCTGCTCG-3′; tln1 rev, 5′-CGCCCTCTTGACAGCATTAC-3′; cyclinb1 (control gene maternally provided) fwd, 5′-TCCATGTTCCTCCGTCTCTC-3′; cyclinb1 rev, 5′-CATGTGCATCTGCTTCTGGT-3′; C18H16orf7 (maternally provided, but not transcribed after zygotic transition) fwd, 5′-TGTCCCATCTCTCCACATCA-3′; C18H16orf7 rev, 5′-GTGAGAAGGAACCCAGTCCA-3′; tal1 (not maternally provided) fwd, 5′-ATGGCTCCATGCACACACTA-3′; tal1 rev, 5′- GTTTCCTTGGCAACACCATT-3′.
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