Open Access
Table 1
Primer sequences for RAA-Cas13a-LFD assay.
| Primers | Sequences (5′ to 3′) | Amplicon sizes (bp) | Description |
|---|---|---|---|
| TOX-F1 | ACTACAGACGCGATGCCGCTCCTCC | 283 | Candidates for RAA primers screened in this study. TOX-F1/TOX-R1 pair was used for follow-up experimental tests. |
| TOX-R1 | TGTCTCCCTCGCCCTCTTCTCCACT | ||
| TOX-F2 | ACACCGGAATGCGATCCAGACGAGA | 126 | |
| TOX-R2 | GTCCAAGCCTCCGACTCTGTCTCCC | ||
| TOX-F3 | ATATCAGGACTGTAGATGAAGGCGAGGGT | 176 | |
| TOX-R3 | CGTCTCGTCTGGATCGCATTCCGGTGTCT | ||
| TOX-F4 | TAGATGAAGGCGAGGGTGAGGATGAGGGG | 121 | |
| TOX-R4 | CTCCAGGAAAAGCAGCCAAGCCGGAAACA | ||
| TOX-F5 | AAGATGTTTCCGGCTTGGCTGCTTTTCCTG | 162 | |
| TOX-R5 | CCGACTCTGTCTCCCTCGCCCTCTTCTCCA | ||
| T7 promoter | GAAATTAATACGACTCACTATAGGG | Promoting the transcription of amplified DNA to RNA. | |
| Probe 1 | CGUCUCGUCUGGAUCGCAUUCCGGUGUC | Expected sequences of guide RNA obtained from in vitro transcription. Probel was used for follow-up experimental tests. | |
| Probe 2–5-a | UUCUCUCCGCCAUCACCACGAGGAAAGC | ||
| Probe 2–5-b | CAAUUCUCUCCGCCAUCACCACGAGGAA | ||
| Probe 3–4-a | CUCCGACUCUCGUCGCUUCCCAACCACG | ||
| Probe 3–4-b | CCGACUCUCGUCGCUUCCCAACCACGCC | ||
| Lateral flow reporter | FAM/mArArUrGrGrCmAmArArUrGrGrCmA/Bio | Trans-cleavage reporter for Cas13a. |
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