FIGURE

Fig. S17

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ZDB-FIG-180126-57
Publication
Ma et al., 2017 - A novel inducible mutagenesis screen enables to isolate and clone both embryonic and adult zebrafish mutants
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Fig. S17

Segregation analysis of causative mutation from the multiple-inserted lines with Dox-dependent fin regeneration defects. (A) Segregation of galnt2IM from line pIDM-A28. To segregate the causative mutation, pIDM-A28 F1 mutants with Dox-dependent fin regeneration defects were crossed with WT. The cross containing about 50% EGFP positive offspring was selected to produce F2 population. Left panel: Southern blot analysis of insertion copy numbers in the pool of embryos as indicated. Pool of EGFP positive F2: The F2 EGFP positive embryos were from the cross between the selected F1 fish and WT; Pool of EGFP positive F3: The F3 EGFP positive embryos were from the cross between one of F2 adult fish and WT. Southern blot was performed as described in Supplementary Figure 3. Right panel: PCR analysis to identify the insertion site. A pair of galnt2 intron 1 and pIDM specific primers were designed to amplify the insertion site. The DNA lysed from caudal fin of individual F2 fish was used to do PCR analysis. The DNA from EGFP positive F1 embryo pool was used as the positive control. The DNA from EGFP negative F2 fish was used as the negative control. (B) Induction of the listed genes in WT zebrafish at different time points during fin regeneration as indicated. The WT caudal fin was sampled at different times after amputation and subjected to a WISH assay with the corresponding anti-sense RNA probes as indicated. (C) Segregation of ggt7lIM from line pIDM-A3. To segregate the causative mutation, pIDM-A3 F2 mutants with Dox-dependent fin regeneration defects were crossed with WT. The cross containing about 50% EGFP positive offspring were selected to produce F3 population. Left panel: Southern blot analysis of insertion copy numbers in the pool of embryos as indicated. Pool of EGFP positive F3: The F3 EGFP positive embryos were from the cross between the selected F2 fish and WT; Pool of EGFP positive F4: The F4 EGFP positive embryos were from the cross between one of F3 adult fish and WT. Southern blot was performed as described in Supplementary Figure 3. Right panel: PCR analysis to identify the insertion site. A primer from ggt7l exon 5 was used with pIDM specific primer to amplify the insertion site. The DNA lysed from caudal fin of individual F3 fish was used to do PCR analysis. The DNA from EGFP positive F1 embryo pool was used as the positive control. The DNA from EGFP negative F3 fish was used as the negative control. (D) Segregation of cry61IM from line pIDM-E7. To segregate the causative mutation, pIDM-E7 F2 mutants with Dox-dependent fin regeneration defects were crossed with WT. The cross containing about 50% EGFP positive offspring were selected to set up F3 population. Left panel: Southern blot analysis of insertion copy numbers in the pool of embryos as indicated. Pool of EGFP positive F3: The F3 EGFP positive embryos were from the cross between the selected F2 fish and WT; Pool of EGFP positive F4: The F4 EGFP positive embryos were from the cross between one of F3 adult fish and WT. Southern blot was performed as described in Supplementary Figure 3. Right panel: PCR analysis to identify the insertion site. A primer from cry61 intron 1 was used with pIDM specific primer to amplify the insertion site. The DNA lysed from caudal fin of individual F3 fish was used to do PCR analysis. The DNA from EGFP positive F1 embryo pool was used as the positive control. The DNA from EGFP negative F3 fish was used as the negative control.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
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