Table 1.

Primers used in experimental procedures

PrimerSequenceDescription
1 AAAAATAATACGACTCACTATAGGTGGGTCTGCAGCTGTA 5′ gRNA amplification primer (polyA, T7, target sequence, target underlined) 
2 GGTGGGTCTGCAGCTGTAGTTTTAGAGCTAGAAAT Target sequence, 5′ pDR274 primer (target underlined) 
3 AAAAGCACCGACTCGGTGCC 3′ pDR274 primer 
4 TTGACAACACGTCACTCCTTG 5′ genotyping primer 
5 TTCTTGCAGATCAGCATTGG 3′ genotyping primer 
6 CAGGCTGGACCAAAACAGACAGAACGTTTCTACC cDNA amplification primer (vector homology underlined) 
7 GCTCCGCTTCCATAGCATTCGAAATCACAACCC cDNA amplification primer (vector homology underlined) 
8 GGAACATGGCAGATCATCCT 5′ primer for in situ probe 
9 GGTAATACGACTCACTATAGcccagcaggtgccagctgac 3′ primer for in situ probe with T7 promoter 
10 TCCCCTAAAAATGCATGTCTGT 5′ intron based genotyping primer 
11 CAAGGCATCTCAGCATGAACA 3′ intron based genotyping primer 
12 GCAGATCATCCTTGCcGCCTCCACCCtCATGGCATTGC p.S83R (D rerio 102) 5′ primer 
13 GCAATGCCATGaGGGTGGAGGCgGCAAGGATGATCTGC p.S83R (D rerio 102) 3′ primer 
14 CGGGAAGCAGTCAGAGGacTCTTTATACTTTGACAACACG p.G97D (D rerio 116) 5′ primer 
15 CGTGTTGTCAAAGTATAAAGAgtCCTCTGACTGCTTCCCG p.G97D (D rerio 116) 3′ primer 
16 GCCGAACTTGGACCTgCTACTCcGCAGTGAATCCTGAGAG p.Y1702C (D rerio 1596) 5′ primer 
17 CTCTCAGGATTCACTGCgGAGTAGcAGGTCCAAGTTCGGC p.Y1702C (D rerio 1596) 3′ primer 
18 GGCATCCTTGGTATGCatGCCTAAATAAACAAGGAACCGC p.R2074C (D rerio 1965) 5′ primer 
19 GCGGTTCCTTGTTTATTTAGGCatGCATACCAAGGATGCC p.R2074C (D rerio 1965) 3′ primer 
20 GCAGAAATCCGTCACCATGtGtATTGAGCTcCTGGGTTGTG p.R2187C (D rerio 2092) 5′ primer 
21 CACAACCCAGgAGCTCAATaCaCATGGTGACGGATTTCTGC p.R2187C (D rerio 2092) 3′ primer 
22 CGGAAGATAAGCAGAGGAAGAG) qPCR forward primer 
23 AGAGCTTGGAATTCTTGGCCCAGT qPCR probe 
24 ACTATAGGGACGTGATGCTTTG qPCR reverse primer 
PrimerSequenceDescription
1 AAAAATAATACGACTCACTATAGGTGGGTCTGCAGCTGTA 5′ gRNA amplification primer (polyA, T7, target sequence, target underlined) 
2 GGTGGGTCTGCAGCTGTAGTTTTAGAGCTAGAAAT Target sequence, 5′ pDR274 primer (target underlined) 
3 AAAAGCACCGACTCGGTGCC 3′ pDR274 primer 
4 TTGACAACACGTCACTCCTTG 5′ genotyping primer 
5 TTCTTGCAGATCAGCATTGG 3′ genotyping primer 
6 CAGGCTGGACCAAAACAGACAGAACGTTTCTACC cDNA amplification primer (vector homology underlined) 
7 GCTCCGCTTCCATAGCATTCGAAATCACAACCC cDNA amplification primer (vector homology underlined) 
8 GGAACATGGCAGATCATCCT 5′ primer for in situ probe 
9 GGTAATACGACTCACTATAGcccagcaggtgccagctgac 3′ primer for in situ probe with T7 promoter 
10 TCCCCTAAAAATGCATGTCTGT 5′ intron based genotyping primer 
11 CAAGGCATCTCAGCATGAACA 3′ intron based genotyping primer 
12 GCAGATCATCCTTGCcGCCTCCACCCtCATGGCATTGC p.S83R (D rerio 102) 5′ primer 
13 GCAATGCCATGaGGGTGGAGGCgGCAAGGATGATCTGC p.S83R (D rerio 102) 3′ primer 
14 CGGGAAGCAGTCAGAGGacTCTTTATACTTTGACAACACG p.G97D (D rerio 116) 5′ primer 
15 CGTGTTGTCAAAGTATAAAGAgtCCTCTGACTGCTTCCCG p.G97D (D rerio 116) 3′ primer 
16 GCCGAACTTGGACCTgCTACTCcGCAGTGAATCCTGAGAG p.Y1702C (D rerio 1596) 5′ primer 
17 CTCTCAGGATTCACTGCgGAGTAGcAGGTCCAAGTTCGGC p.Y1702C (D rerio 1596) 3′ primer 
18 GGCATCCTTGGTATGCatGCCTAAATAAACAAGGAACCGC p.R2074C (D rerio 1965) 5′ primer 
19 GCGGTTCCTTGTTTATTTAGGCatGCATACCAAGGATGCC p.R2074C (D rerio 1965) 3′ primer 
20 GCAGAAATCCGTCACCATGtGtATTGAGCTcCTGGGTTGTG p.R2187C (D rerio 2092) 5′ primer 
21 CACAACCCAGgAGCTCAATaCaCATGGTGACGGATTTCTGC p.R2187C (D rerio 2092) 3′ primer 
22 CGGAAGATAAGCAGAGGAAGAG) qPCR forward primer 
23 AGAGCTTGGAATTCTTGGCCCAGT qPCR probe 
24 ACTATAGGGACGTGATGCTTTG qPCR reverse primer 

qPCR, quantitative PCR.

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