Table 1.
Locus | Primer sequences (5′–3′) | Repeat motif | Ta (°C) | Ct (µM) | Multiplex | Fluorescent dye | Post-PCR dilution | Position A.t.a | Position N.c.a | Chromosomal blocksb | Allele size range (bp) | GenBank accession no. | Publication |
Ncpm09 | F: TAGACGCTGCGTTTTGAAGA | CT18 | 58 | 0.4 | NcM1 | 6-FAM | 1/150 | 1 | 2 | B | 96–130 | KR065729 | |
R: CCTCTGTTGAGTGAATGGTTCTC | |||||||||||||
Ncpm13 | F: CCAAAACTATGCCGATCTCA | AG16 | 58 | 0.2 | NcM1 | VIC | 1/150 | NA | NA | NA | 158–166 | KR065730 | |
R: CCACGAGCGAATCTTCTTGT | |||||||||||||
Ncpm21 | F: GTCACCACTTGCTACGGGAT | CTT10 | 58 | 0.2 | NcM1 | VIC | 1/150 | 3 | 7 | F | 240–277 | KR065731 | |
R: CATTGGATAGCACAGAGGCA | |||||||||||||
Ncpm23 | F: TTTCATGTCTCGGATCCTCC | TTC11 | 58 | 0.4 | NcM1 | 6-FAM | 1/150 | NA | NA | NA | 197–263 | KR065732 | |
R: GCAGAGCGCAATCTAAGGAC | |||||||||||||
Ncpm31 | F: GATTATCGAGCTTACTAAAAGCAGC | CTT12 | 58 | 0.2 | NcM1 | NED | 1/150 | 5 | 4 | W | 58–101 | KR065733 | |
R: GTGTTGAAGCGCAATGAAGA | |||||||||||||
Tc-up1 | F: TGCTCTGTTTCTCTCCACATTC | (CA)n(CT)n | 50 | 0.2d | NcM1 | NED | 1/160 | NA | NA | NA | 132–170 | AJ746212 | Basic and Besnard, 2006 |
R: TTCCTTGCTTCTTCTCTTCCA | |||||||||||||
Ncpm07 | F: TGGAATGGTTCTGTGGACAA | TCA18 | 58 | 0.4 | NcM2 | 6-FAM | 1/150 | NA | NA | NA | 109–139 | KR065734 | |
R: TTCTGGAATTGGCTGCTTCT | |||||||||||||
Ncpm14 | F: GACCACATCTCGTCTTGCCT | CTT15 | 58 | 0.4 | NcM2 | PET | 1/150 | NA | NA | NA | 108–123 | KR065735 | |
R: GACCCTAACTACAGGCTGTGAAA | |||||||||||||
Ncpm19 | F: CCAACAATGGATTGGGAGAG | GTGTA(TG)14 | 58 | 0.4 | NcM2 | VIC | 1/150 | NA | NA | NA | 100–124 | KR065736 | |
R: TCCCCATCACTCCAGCTAAG | |||||||||||||
Ncpm29 | F: CTCCCTCTTCTCCTACCTACACATA | AC17 | 58 | 0.4 | NcM2 | NED | 1/150 | NA | NA | NA | 78–94 | KR065737 | |
R: GGTTGAAAGTAGGAGTGAGTCAAGA | |||||||||||||
Tc-up4 | F: GTTTTGTCCGCTTTGCTTCC | CT13 | 50 | 0.2d | NcM2 | VIC | 1/140 | NA | NA | NA | 253–262 | AJ746216.1 | Basic and Besnard, 2006 |
R: GCCATAGACTTTCTCATTGATTC | |||||||||||||
9C7/Thlc3 | F: GTCACGAGTTTCACCATT | AG13 | 58 | 0.4 | NcM2 | VIC | 1/150 | NA | NA | NA | 152–191 | Jiménez-Ambriz et al., 2007 | |
R: ATCTTCCACAATTGTGCC | |||||||||||||
Tc-up2 | F: TGAGAAGAGGAGACACAGGAAC | (AG)5(AG)5(GA)6 | 58 | 0.2 | NcM2 | PET | 1/150 | NA | NA | NA | 232–244 | AJ746213.1 | Basic and Besnard, 2006 |
R: CACTTACCAAATCGAAAACTGCTCC | |||||||||||||
Nc02 | F: GGAGCTGTGGTTTCTGAAGG | AGG8 | 68/47c | 0.06 | NcM3 | VIC | None | 4 | 3 | O | 170–191 | KR065738 | |
R: AGCATCGTATTCCGATCCAG | 0.28 | ||||||||||||
Nc03 | F: TGAGCTTCTTCAGTCCCGAT | AC12 | 68/47c | 0.06 | NcM3 | NED | None | 5 | 4 | S | 188–194 | KR065739 | |
R: TATGAGCTCGTCGCTCACAG | 0.28 | ||||||||||||
Nc04 | F: ACGGTCGCATACCAAAAAGT | AG11 | 68/47c | 0.06 | NcM3 | VIC | None | 2 | 5 | J | 123–143 | KR065740 | |
R: AGGATGCACTCCTTGAGACC | 0.28 | ||||||||||||
Nc06b | F: GCGTCTTCTCTCCATCCTCA | AG13 | 68/47c | 0.07 | NcM3 | PET | None | 4 | 6 | U | 89–153 | KR065741 | |
R: GGATTTCCAATTCAATCTTCCC | 0.37 | ||||||||||||
Nc07b | F: CCAGTTTCCAACGGCATAGT | AC10 | 68/47c | 0.06 | NcM3 | NED | None | 3 | 7 | F | 105–112 | KR065742 | |
R: TTGGTTTGGTTTCTTCTTGTGA | 0.28 | ||||||||||||
Nc19 | F: CGGATTGTTGTGAATCCCAT | AGG11 | 68/47c | 0.07 | NcM3 | PET | None | 2 | 5 | J | 199–235 | KR065743 | |
R: ACCTCTTCTTTCGCCCTTGT | 0.28 | ||||||||||||
Nc20 | F: ACACAACTCCAAAGGCTTCA | AG11 | 68/47c | 0.15 | NcM3 | 6-FAM | None | 4 | 3 | O | 205–234 | KR065744 | |
R: TTTTGTTCTAACCGTTACTCTTT | 0.75 | ||||||||||||
Nc22 | F: TTGCTTTCACATGTCTTGACG | AG10 | 68/47c | 0.11 | NcM3 | 6-FAM | None | 2 | 5 | K | 111–121 | KR065745 | |
R: GAACAGAACAAGAAGACATGAATGA | 0.56 |
Note: Ct = final concentration of primers; NA = not available; Ta = annealing temperature.
Relative chromosomal position in Arabidopsis thaliana and Noccaea caerulescens, respectively (Schranz et al., 2006).
Genome blocks in the “ancestral karyotype” blocks (Schranz et al., 2006).
Following Godé et al. (2012), touchdown PCR was performed. During the first 10 cycles, annealing temperature was decreased from 68°C to 50°C in increments of 2°C. This was followed by 27 cycles with an annealing temperature at 47°C.
Standalone PCR.