Table 1. Markers developed for dreissenid identification derived from MPS output and qPCR conduction.
| Single-copy gene calling step |
PCR amplification step |
qPCR amplification step |
GenBank accession | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| MPS Contig reference | BlastX E- value | Predicted gene | Primer sequences (5′ → 3′) | Size (bp) | Ta (°C) | Tm (°C) | E | CV | R2 | DL (ng/μl) | QL (ng/μl) | |
| Contig000070 | 3.77E-88* | H3 | F: GGTGACACGCTTGGCGTGGA | 229 | 60 | — | — | — | — | — | — | JWHF01000070 |
| R: GCCAGGAACCGTCGCCCTTC | ||||||||||||
| Contig000076 | 5.31E-54* | H2B | F: CGCGCGCTCCACTGACAAGA | 251 | 60 | 85.4692 ± 0.2057 | 1.9729 | 0.1541 | 0.9976 | 5E-4 | 5E-4 | JWHF01000076 |
| R: CACCAGGCAGCAGGAGACGC | ||||||||||||
| Contig000102 | 1.08E-27 | H1 | F: TCTTGGCGCCCGCCTTCTTG | 214 | 60 | 86.0933 ± 0.2612 | 1.9700 | 0.2283 | 0.9954 | 5E-3 | 5E-3 | JWHF01000102 |
| R: GTCAGTGCCGTCAACGCCCA | ||||||||||||
| Contig000913 | 7.52E-23 | MARS | F: AGTCCTCCCAGATTAGCCTGTGC | 277 | 65 | 80.8778 ± 2.6423 | 1.9102 | 0.1772 | 0.9940 | 5E-2 | 5E-1 | JWHF01000913 |
| R: AGATGTCGCGGTGGAGGGCT | ||||||||||||
MPS contig reference, Blast Result, predicted gene, Forward (F) and Reverse (R) primer sequences, amplicon size in base pairs, and optimum annealing temperature (Ta) for PCR amplification, Melting temperature (Tm) in Real Time PCR, efficiency (E), coefficient of variation (CV), coefficient of regression (R2), detection (DL) and quantification (QL) levels and GenBank accession number for four single-copy predicted genes selected for zebra mussel. *Significant BlastX E-value <1E-50.