Fig. 2.

Optimization of PCR tagging injection mix. (A,B) Optimization of homology arm (HA) length, template concentration and end modification (A) and small molecule modulation of NHEJ/HDR pathways (B). Efficiency is expressed as percentage of F0 larvae that are mCherry⁺ or mEGFP⁺ at 3-5 dpf. Circles indicate individual injection rounds and bars indicate the mean of 3-5 injection rounds per condition. The number of surviving larvae analyzed per injection round varied between 9 and 253 (88 on average, for full details see Table S2). Error bars indicate standard deviation. *P<0.05, **P<0.01, ***P<0.001 (two-way ANOVA corrected for multiple comparisons). For ease of comparison, data indicated by superscript a and b are shown in more than one graph. (C) Suggested components of injection mix for PCR tagging in zebrafish. Biosg, standard biotin modification; PS, phosphorothioate bond.