Table 1 Candidate reference genes, amplicon characteristics and primer sequences
Gene | Locus name in Oat Transcriptome [1] | RPKM [1] | Primers (5′–3′) | Tm (℃) | Amplicon length | Efficiency (%) | R2 |
|---|---|---|---|---|---|---|---|
PP2A | Locus_1956_Transcript_6/10_Confidence_0.500_Length_1648 | 27.3738 | F: GCCCTGAGCCTACAAGAACGG | 63.6 | 196 bp | 100.4 | 1 |
R: GCTGAGCGAACATGCTGAGAT | 60.3 | ||||||
UBQ10 | Locus_4160_Transcript_1/1_Confidence_1.000_Length_1516 | 12.1493 | F: GATCTCAGCTCTAGCGAATCTCC | 59.4 | 136 bp | 94.2 | 1 |
R: ATCCCTTCCTTGTCTTGAATCTTG | 60.9 | ||||||
EF1A | Locus_565_Transcript_5/10_Confidence_0.529_Length_863 | 347.2181 | F: GTGAAGATGATTCCCACCAAGC | 60.9 | 87 bp | 96.0 | 1 |
R: CCTCATGTCACGCACAGCAAA | 62.7 | ||||||
HNR | Locus_4951_Transcript_1/4_Confidence_0.667_Length_1455 | 12.2775 | F: ATTGGGTTTGTCACTTTCCGTAG | 60.2 | 134 bp | 101.5 | 1.000 |
R: CTTGGAGGGTGTCTCGCATCT | 61.4 | ||||||
EP | Locus_827_Transcript_1/2_Confidence_1.000_Length_1359 | 19.8139 | F: GCACAAGTGATGCCAGAATAGC | 60.0 | 193 bp | 112.4 | 1 |
R: CGAGATGCATTAGATTCGTTGG | 60.1 | ||||||
TBC | Locus_9878_Transcript_1/1_Confidence_1.000_Length_1218 | 4.3017 | F: TCCTCTTTCACCTCCCGATTAC | 60.0 | 98 bp | 92.7 | 1 |
R: CAGATGCTTGCCCTTCTACCTC | 60.7 | ||||||
EIF4A | Locus_3892_Transcript_3/4_Confidence_0.667_Length_1160 | 45.0042 | F: TCTCGCAGGATACGGATGTCG | 63.3 | 88 bp | 100.8 | 0.99 |
R: TCCATCGCATTGGTCGCTCT | 63.6 | ||||||
18S | Locus_29_Transcript_3/10_Confidence_0.100_Length_1865 | 1067.4606 | F: TTCTTAGTTGGTGGAGCGATTT | 58.7 | 150 bp | 100.4 | 1 |
Locus_29_Transcript_4/10_Confidence_0.100_Length_1865 | 1052.7564 | R: CCTGTTATTGCCTCAAACTTCC | 58.5 | ||||
GAPDH1 | Locus_535_Transcript_3/4_Confidence_0.500_Length_1198 | 291.0028 | F: CTTCAACATCATTCCCAGCAG | 58.0 | 288 bp | 110.7 | 1 |
Locus_1038_Transcript_1/4_Confidence_0.583_Length_1013 | 393.7014 | R: GCCTTGGCGTCAAAGATGCT | 62.6 | ||||
TUA6 | Locus_855_Transcript_1/4_Confidence_0.727_Length_1544 | 28.0927 | F: CCCAACAATGTGAAGTCCAGC | 60.2 | 121 bp | - | - |
Locus_882_Transcript_1/5_Confidence_0.692_Length_1361 | 164.8515 | R: TGAACTGCTCACTCACCCTCC | 59.7 | ||||
Locus_3988_Transcript_3/5_Confidence_0.615_Length_1286 | 24.3489 | ||||||
UBC21 | Locus_10069_Transcript_1/3_Confidence_0.750_Length_710 | 7.3384 | F: GCCCATCGGAGACACCTTTTG | 64.0 | 133 bp | 97.8 | 0.99 |
Locus_10069_Transcript_2/3_Confidence_0.750_Length_723 | 6.3587 | R: CCTGTCTTGAAGTGAACATTTGG | 59.1 | ||||
Locus_3507_Transcript_3/4_Confidence_0.500_Length_967 | 13.6740 | ||||||
Locus_3507_Transcript_2/4_Confidence_0.700_Length_1018 | 16.8311 |