From: A high-throughput method for the detection of homoeologous gene deletions in hexaploid wheat
gene name | sequence length (bp)1 | GenBank accessions | primer sequences2 ; amplicon size1 | probe sequences2 | assay conditions3 | samples screened; mutations detected |
---|---|---|---|---|---|---|
TaNFXL1 | ~630 | HQ595068 - HQ595070 | FNFXL1gsprb GGCACACCAACTCCATCATG RNFXL1gsprb GGCTGGTTGCATCTGATATCCT Amplicon size: ~170 bp | NFXL1prb1FAM AACACATTCTCTTGTGACTG NFXL1prb2VIC AACGCACTCTCTCATGA NFXL1prb3NED ACATTCTCTCATTACTGACACA | FR primers: 1000 nM each Probes: 50 nM FAM, 100 nM VIC, 100 nM NED 20 μL reaction: 18 μL stock; 2 μL DNA sample 3-step reaction: 95°C 15 s denaturation, 55°C 30 s annealing, 60°C 30 s extension - 45 cycles | 2772; 14 |
TaCesA8 | ~370 | HQ595065 - HQ595067 | FCesA8gsprb CCACATGGCGAAGAACACCT RCesA8gsprb AGTGGACGACGGTGCTGATC Amplicon size: ~140 bp | CesA8prb1FAMCTCATACCCGCTATTGA CesA8prb2VICCCGACCAGGTTGAG CesA8prb3NED ACGAGGTTGAGAACC | FR primers: 1000 nM each Probes: 50 nM FAM, 100 nM VIC, 100 nM NED 20 μL reaction: 18 μL stock; 2 μL DNA sample 3-step reaction: 95°C 15 s denaturation, 55°C 30 s annealing, 60°C 30 s extension - 45 cycles | 3762; 19 |
TaPLDβ1 | ~620 | HQ595071 - HQ595073 | FPLDB1gsprb TTTGTACTTGTCTTCCTGATGTTATGC R1PLDB1gsprb GGCGTAGTTGGAGGCAGTCTC R2.3PLDB1gsprb GGTGTAGTTGGAGGCAGTCTCTC Amplicon size: ~100 bp | PLDB1prb1FAM TTGCTTAGAACTGCTGC PLDB1prb2NED CTTAGAACCGCTGCTT PLDB1prb3VIC TAGAACCACTGCTTC | FR primers: 1000 nM F primer, 666 nM R2.3 primer, 333 nM R1 primer Probes: 50 nM FAM, 100 nM VIC, 150 nM NED 11 μL reaction: 9 μL stock; 2 μL DNA sample 3-step reaction: 95°C 15 s denaturation, 55°C 30 s annealing, 60°C 30 s extension - 45 cycles | 3366; 11 |