TABLE 1 .
Primers used to create single guide RNAs (sgRNAs), a donor plasmid, and a qPCR plasmid standard
| Primer name | Sequence (5'-3') | Description |
|---|---|---|
| sgRNA Forward | CACCGTTGGAATGTGGACTTAAGGA | sgRNA to US6 stop codon |
| sgRNA Reverse | AAACTCCTTAAGTCCACATTCCAAC | sgRNA to US6 stop codon |
| Primer 1 | CGGCCCAATTTAATCAAGG | US6 homology arm, forward |
| Primer 2 | AGGATGGTGAGTTGTATGTA | US6 homology arm, reverse |
| Primer 3 | GTCCACATTCCAATCGAGTT | US7 homology arm, forward |
| Primer 4 | AACACCGAAAGGCCAAATAC | US7 homology arm, reverse |
| Primer 5 | ATGGATAGCACTGAGAACGT | DsRed Express2, forward |
| Primer 6 | TTACTGGAACAGGTGGTGGC | DsRed Express2, reverse |
| Primer 7 | ACTCACCATCCTATGGATAGCACT | US6/DsRed overhang, forward |
| Primer 8 | TGGAATGTGGACTTACTGGAACAG | US7/DsRed overhang, reverse |
| Primer 9 | AGTGCTATCCATAGGATGGTGAGT | US6/DsRed overhang, reverse |
| Primer 10 | CTGTTCCAGTAAGTCCACATTCCA | US7/DsRed overhang, forward |
| US7 plasmid standard forward | CTTTCCGGTCCTGTCTCCAC | qPCR forward primer |
| US7 plasmid standard reverse | GGTTAAATCTTACCCGCAGTGC | qPCR reverse primer |