Table 2. Details of primers.
| Target Liposcelis species | Primer Name | Primer Sequence (5’-3’) | Length (bp) | GC% | *Tm | **ΔG | ^ any | ^^ 3’ | Intended use |
|---|---|---|---|---|---|---|---|---|---|
| Liposcelis brunnea | BruCo5F | GGGGTTTTAGGGTTTGTGGTA | 21 | 47.2 | 60.0 | 0.8 | 2 | 2 | Endpoint/qPCR multiplex |
| BruCo5R | AATGTCGCCAACCATCTAAAG | 21 | 42.9 | 59.1 | 0 | 4 | 2 | Endpoint/qPCR multiplex | |
| BruCo6F | TTTATGCGATAGGAGCGATTG | 21 | 42.9 | 60.2 | 0.5 | 4 | 2 | Single PCR | |
| BruCo6R | CATCCAACCCGACAGTAAACA | 21 | 47.7 | 60.7 | 1.0 | 3 | 0 | Single PCR | |
| Liposcelis bostrychophila | BosCo7F | CGATCCCTACCGGAGTTAAAG | 21 | 52.4 | 60.0 | 0.7 | 4 | 1 | Endpoint PCR multiplex |
| BosCo7R | TGGGCAACAACATAGTATCTATCG | 24 | 41.7 | 60.3 | 0.7 | 6 | 4 | Endpoint PCR multiplex | |
| BosCo8F | ATGCTCTCAATCGGAGCTCTAG | 22 | 50.0 | 60.1 | 0.7 | 6 | 4 | qPCR multiplex | |
| BosCo8R | ACTTTAACTCCGGTAGGGATCG | 22 | 50.0 | 60.7 | 0.9 | 4 | 2 | qPCR multiplex | |
| BosCo9F | TGGGCTAATCTCTCACATCATCT | 23 | 43.5 | 60.1 | 0.9 | 3 | 3 | Single PCR | |
| Liposcelis decolor | DecCo10F | CCGGCTTTTGGTATTATTTCAC | 22 | 40.9 | 59.8 | 0.7 | 4 | 1 | Single PCR |
| DecCo10R | ATTATCCCCATTACCCCAAA | 20 | 40.0 | 58.0 | 0.9 | 3 | 0 | Single PCR | |
| DecCo11F | CGAGCTTATTTTACTTCTGCGACT | 24 | 41.7 | 60.4 | 0.9 | 4 | 1 | Endpoint/qPCR multiplex | |
| DecCo11R | TGATCCATACAACGTAGCTAGTCA | 24 | 41.7 | 58.9 | 1.0 | 6 | 2 | Endpoint/qPCR multiplex | |
| Liposcelis obscura | ObsCo12F | GGACAGGGTGGACGGTTTATC | 21 | 57.1 | 62.8 | 1.0 | 3 | 1 | qPCR multiplex |
| ObsCo12R | CTGATTCCTGCTAAATGAAGAGAG | 24 | 41.7 | 58.8 | 0.9 | 3 | 0 | qPCR multiplex | |
| ObsCo13F | AGCTATTGCTCACGGAGGATA | 21 | 47.6 | 59.0 | 0 | 6 | 4 | Endpoint PCR multiplex | |
| ObsCo13R | CCAATTGCGGACATAGCATAA | 21 | 42.9 | 60.8 | 1.0 | 6 | 1 | Endpoint PCR multiplex | |
| Liposcelis pearmani | PeaCo14F | CTGACTTTTTCCCCCTTCACT | 21 | 47.6 | 59.6 | 0.9 | 3 | 1 | qPCR multiplex |
| PeaCo14R | GCCAGGGTGTGAGATTCTAAA | 21 | 47.6 | 59.2 | 0.9 | 5 | 3 | Endpoint/qPCR multiplex | |
| PeaCo15F | TGGTGTGTGAGCAGGTATGGT | 21 | 52.4 | 61.5 | 0.8 | 2 | 0 | Endpoint PCR multiplex |
Sequences of Liposcelis species specific primers and their thermodynamic features used for endpoint PCR and real-time TaqMan qPCR.
*The melting temperature of the primer calculated using Primer 3;
**Plot ΔG value in plot calculated by mFOLD;
^ The self-complementarity score of the oligo (tendency of oligo to anneal to itself or form a secondary structure) calculated using Primer 3;
^^3’ self-complementarity of the oligo (tendency to form a primer-dimer with itself) calculated using Primer3.