Figure 4—figure supplement 2—source data 4.

Figure 4—figure supplement 2—source data 4. CRISPR guide RNAs against trib1 and trib3 efficiently disrupt a restriction digest site. (A) Electrophoresis gel of larvae injected with trib1 guide RNA with a SacII restriction digest. The undigested DNA band, obtained using the polymerase chain reaction (PCR), is 177 bp and the successfully digested band is 132 bp. Two uninjected fish were used as controls without (U) and with (D) digestion. Four tyrosinase (tyr) control guide injected larvae had conserved SacII sites while 6/6 trib1 larvae did not digest with SacII indicating a genomic disruption at the CRISPR PAM site. First and last lanes of gel images contain Low Molecular Weight DNA Ladder (New England Biolabs, N3233) for reference. (B) Electrophoresis gel of larvae injected with trib3 guide RNA with a MwoI restriction digest. The undigested PCR band is 186 bp and the successfully digested band is 103 bp. Two uninjected fish were used as controls without (U) and with (D) digestion. Four tyrosinase (tyr) guide injected larvae had conserved MwoI sites while 5/6 trib3 larvae did not digest with MwoI indicating a genomic disruption at the CRISPR PAM site. First and last lanes of gel images contain NEB Low Molecular Weight ladder for reference.