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Fig. 8 Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of wnt9a (top panel) in median fins of zebrafish (5.6-6.3 mm). Columns 1 and 2 contain 1-kb and 100-bp ladders, respectively, with 1-kb and 500-bp bands marked at left. Column 9 shows RT-PCR results with no template added to the reaction mix (negative control). Columns 6-8, with 36, 50, and 76 hours postfertilization (hpf) whole embryos, and the bottom panel, with constitutively expressed ef-1α (Nordnes et al.,[1994]), were used as positive controls. Columns 3-5 contain the RT-PCR reaction product for caudal fin rays, caudal hypurals, and anal fin tissue. Note the absence of wnt9a RNA in both caudal fin rays (column 3) and caudal hypurals (column 4), and lack of amplification of the expected ∼440-bp fragment in anal fin tissue (column 5) that is observed in 36, 50, and 76 hpf embryos. Lanes 5-8 exhibit several bands between 1 and 1.5 kb. These could represent tissue-specific splice variants or nonspecific primer binding. No additional ef- 1α bands were observed (not shown).