Figure 3: Gene expression analysis of endogenous zNMT during embryogenesis. (A) Total RNA was prepared from embryos at 6 hpf and aliquots of 30 μg were separated on agarose gel containing formaldehyde. The RNA were then transferred to nylon membrane by capillary blotting, hybridized with DIG-labeled RNA probes which hybridize specifically to zNMT1 and zNMT2 mRNA. The hybridized probes were detected with AP conjugated anti-DIG antibody and BM purple AP substrate. (B) Total RNA was prepared from the embryos at indicated stages and cDNAs were reverse-transcribed. 30 cycles of PCR for zNMT1, zNMT2 and actin 2 were performed, amplification of those cDNA were analyzed by agarose gel electrophoresis followed by EtBr staining.