Table 1 Primers and Probes employed for LAMP-CRISPR and Real-time PCR assay
From: An isothermal CRISPR- based lateral flow assay for detection of Neisseria meningitidis
N. meningitidis gene target | Sequences of Primer and Probes (5’-3’) | Tm in °C | |
|---|---|---|---|
LAMP-CRISPR/Cas (Primers, gRNA and probes) | ctrA-F3 | TCCGCGACAAAATATTTTGCTGC | 70 |
ctrA-B3 | CCATTTATCCTGACGTTCTGCCG | 70 | |
ctrA-LF | CACCGCACCCATAGACGTAA | 67 | |
ctrA-LB | CGTGGTGTGTTTGTGTTCCG | 69 | |
ctrA-FIP | ACCGATTTCTTGTGTTCTCCCCATGATTACCAATCCCT | 63 − 60 | |
ctrA-BIP | GTATGGGCGGTTTGCAAGATCGCATTCCACCAATGGCGT | 58–63 | |
ctrA-sgRNA | UAAUUUCUACUAAGUGUAGAUAGCCAGAGGCUUAUCGCUUUC | 62 | |
ssDNA FAM-biotin | /5(6)-FAM/TTATTATT/3BIO/ | - | |
ssDNA FQ reporter | /5(6)-FAM/TTATTATT/3BHQ-1/ | - | |
Real-time PCR | ctrA_F | GCT GCG GTA GGT GGT TCA A | 58 |
ctrA_R | TTG TCG CGG ATT TGC AAC TA | 45 | |
ctrA_probe | /FAM/CAT TGC CAC GTG TCA GCT GCA CAT/TAMRA/ | 54 |