Table 3.
LAMP (loop‐mediated isothermal amplification) primer and amplicon sequences (gBlock) and parameters
| LAMP primer or amplicon | Sequence 5′–3′ | Primer length (bp) | Predicted Tm, annealing temperature (°C) | Degeneracy of primer (fold) |
|---|---|---|---|---|
| Khapra_gBlock fragment | cccGGTAATTTAATCTTATAATCACAAGATGGcccATCATCTAATCATAAATCAATGTTTCAcccTAGTCACCCCAACCAAATTAAcccCCTAAAATTGAAAATTTCTATACTAACAAcccTTTAACAATTAAAGAAATAATAAAACTCTcccCGTCTTTTAAAAAAATTTGAGCCcccCAAAATAAAAAAGAGACAGTAATCAcccTTCGTCCAACCATTCATTCCAGTTccc | 234 | N/A | None |
| Khapra_F3 | GGTAATTTAATCTTATAATCACAAGATGG | 29 | 60.9 | None |
| Khapra_B3 | AACTGGAATGAATGGTTGGACGAA | 24 | 69.2 | None |
| Khapra_FIP | TTGTTAGTATAGAAATTTTCAATTTTAGGATCATCTAATCATAAATCAATGTTTCA | 56 | 73.8 | None |
| Khapra_BIP | TTTAACAATTAAAGAAATAATAAAACTCTTGATTACTGTCTCTTTTTTATTTTG | 54 | 70.7 | None |
| Khapra_Floop | TTAATTTGGTTGGGGTGACTA | 21 | 60.7 | None |
| Khapra_Bloop | CGTCTTTTAAAAAAATTTGAGCC | 23 | 61.6 | None |
| Insect 18S | ||||
| 18S_F3 | AGAGGTGAAATTCTTGGATCGTC | 23 | 64.3 | None |
| 18S_B3 | CCCGTGTTGAGTCAAATTAAG | 21 | 61.0 | None |
| 18S_FIP | GGTTAGAACTAGGGCGGTATCKAAGCGAAAGCATTTGCCA | 40 | 80.1 | 2 |
| 18S‐BIP | TCCGGGGGAAGTATGRTTGCAAAGGCTCCACTCCTGGTGGT | 41 | 88.9 | 2 |
| 18S_Floop | GCCTTCGAACCTCTAACTTTC | 21 | 60.8 | None |
| 18S_Bloop | TGAAACTTAAAGGAATTGACGGAA | 24 | 64.4 | None |
The F2 and B2 primer regions of FIP and BIP are underlined. Lowercase letters in the gBlock indicate extra c's added between LAMP primer sites to increase the overall melting temperature (Tm) of the amplicon.