Table 1. Primer sequences, characteristics, polymorphism levels, and genetic diversity of 21 species-specific microsatellite loci in 88 individuals of Prochilodus magdalenae randomly chosen from the whole sample.
| Locus | Primer sequence (5′−3′) | Motif | Ta (°C) | Na | Ra | PIC | HO | HE | P |
|---|---|---|---|---|---|---|---|---|---|
| Pma39a | F: CCAATGACCTGTTTTCTACATTTGG | (ATCT)n | 58 | 14 | 231–283 | 0.860 | 0.671 | 0.878 | 0.002 |
| R: AATCTACTACCCGGATGGCG | |||||||||
| Pma25a | F: AAGGGGAAAGAAATCCAGGC | (AAGGC)n | 60 | 12 | 174–229 | 0.816 | 0.795 | 0.840 | 0.003 |
| R: ATCCTGGGTTCATACCGACG | |||||||||
| Pma02a | F: CGACATTCAACATGACAGTGC | (ATCT)n | 58 | 19 | 231–307 | 0.917 | 0.816 | 0.927 | 0.019 |
| R: CACCAAATTGATGCAAACTGC | |||||||||
| Pma35a | F: GCAGTCTGGCATTTTAGTGGC | (ATCT)n | 58 | 21 | 269–353 | 0.935 | 0.536 | 0.944 | 0.000 |
| R: ACCACATCTCGCATCACTGG | |||||||||
| Pma56c | F: ATTTGGTGCCTGTAGCTGGG | (ATT)n | 60 | 37 | 132–279 | 0.949 | 0.670 | 0.956 | 0.000 |
| R: ACGGTCGGTGCACTAATTCC | |||||||||
| Pma01a | F: TTGTCATTTCCCGGTTTTCC | (ATCT)n | 58 | 25 | 216–344 | 0.938 | 0.753 | 0.947 | 0.000 |
| R: TGGCCCAGCTGTAATTTGG | |||||||||
| Pma40a | F: CTGGTTACCCACCACTGTCG | (ATCT)n | 58 | 25 | 236–344 | 0.932 | 0.686 | 0.941 | 0.000 |
| R: CACATTGCCATTTGGAGACG | |||||||||
| Pma46a | F: TTGATGTAAACATCTCATTGCCG | (ATCT)n | 56 | 19 | 126–198 | 0.918 | 0.830 | 0.929 | 0.005 |
| R: TTGCTGGAGGTTCTGTCCG | |||||||||
| Pma36a | F: TCATGATGAAATGCCACACC | (ATCT)n | 58 | 24 | 119–219 | 0.925 | 0.674 | 0.935 | 0.000 |
| R: TGCACGTGAACTTAGGCACC | |||||||||
| Pma18a | F: ACTGAGACAAAACCCGGAGG | (ATT)n | 62 | 13 | 209–251 | 0.728 | 0.471 | 0.755 | 0.000 |
| R: CTTCATACACCCACCATCAGG | |||||||||
| Pma13a | F: CCGAAGCTATTTACCCAGCG | (AAAT)n | 62 | 11 | 154–194 | 0.815 | 0.670 | 0.841 | 0.007 |
| R: TGAAATATGCTCGTGCTCCC | |||||||||
| Pma14a | F: GTTCAGGGTCCTGCTGTTCC | (TTC)n | 58 | 21 | 146–209 | 0.907 | 0.605 | 0.919 | 0.000 |
| R: TTTCGGTGTTGGAACATTGC | |||||||||
| Pma42c | F: TTACACAGCGTCCCAATTCC | (ATCT)n | 58 | 25 | 146–254 | 0.933 | 0.759 | 0.942 | 0.000 |
| R: GCTGCAGGGATTGTCCTACC | |||||||||
| Pma26c | F: TGATGTTTCCTCCCCTCACC | (ATCTC)n | 58 | 20 | 141–281 | 0.888 | 0.553 | 0.902 | 0.000 |
| R: GTGTTTCCTGCTCTCTGCCC | |||||||||
| Pma34d,e | F: GAGTGCCGATGACAGAGACG | (ATCT)n | 58 | 24 | 202–406 | 0.919 | 0.363 | 0.930 | 0.000 |
| R: CAAGATGCCCTGTAGTGCCC | |||||||||
| Pma50c | F: GATTCCTTCCTACCGGAGCC | (ATCT)n | 58 | 30 | 171–299 | 0.942 | 0.565 | 0.950 | 0.000 |
| R: ATGAGCACCACCCTCAATCC | |||||||||
| Pma32f | F: GAAAAGACACAACAGCGCCC | (ATCT)n | 58 | 13 | 146–294 | 0.399 | 0.375 | 0.430 | 0.006 |
| R: GTCGCTAATAGCCATGCCG | |||||||||
| Pma57b | F: ATGGCAATGGTTAAGGGTCG | (AAC)n | 58 | 11 | 191–230 | 0.838 | 0.306 | 0.861 | 0.000 |
| R: CTGAAAGCCCCTGTTTGTGC | |||||||||
| Pma08b,e | F: TTTTATTATTCCCCATTTTCTCCC | (AAAG)n | 58 | 12 | 254–298 | 0.833 | 0.257 | 0.856 | 0.000 |
| R: TGGGTTTTGAGCTGTTCTGC | |||||||||
| Pma17b | F: CTGTGGGCAGCAAAGTGC | (ATT)n | 58 | 36 | 151–346 | 0.892 | 0.595 | 0.904 | 0.000 |
| R: CTTTGAGCCACTTCAAACGG | |||||||||
| Pma47b | F: TGGCTGCTAAATTAAATCCTTTGG | (ATCT)n | 58 | 21 | 176–280 | 0.915 | 0.413 | 0.928 | 0.000 |
| R: AAGCAAAACCGTTCCACAGC | |||||||||
| Across loci | 20.619 | 119–353 | 0.867 | 0.589 | 0.876 | 0.000 |
Notes:
Ta: annealing temperature standardized in PCRs, Na: number alleles per locus; Ra: allelic size range (bp); PIC: polymorphism information content; HO and HE: observed and expected heterozygosities, respectively; P: statistical significance (values in bold represent significance at P < 0.05).
a
Satisfied selection criteria.
b
inconsistent amplifications.
c
Low definition peaks.
d
Dropout.
e
Stuttering.
f
Low value of PIC.