From: Validation of a set of reference genes to study response to herbicide stress in grasses
Reference gene1 (accession number) | Primer sequences (5'-3')2 | Amplicon lenght (bp) | Tm (°C) | Manual threshold3 | PCR efficiency (%) | Regression coefficient (R2) | Average Cq value |
---|---|---|---|---|---|---|---|
CYC | F: AGCTTTGAAGTTGGCAGTAG | ||||||
R: GATCGCGTATTCATGGACTTTAG | Discarded (aspecific amplification) | ||||||
SPS | F: CATTGCAAGAACTATTTGTCACG | ||||||
R: GCAGAGATCAAATGGTTCAAATC | |||||||
ACT | F: TGTGCTTGACTCTGGTGATG | 220 | 58 | ||||
R: TTCATAATCAAGGGCAACGTAAGC | Discarded (aspecific amplification) | ||||||
RUBISCO | F: CATTATCAAGAAGGGCAAGATGTG | 169 | 60 | ||||
R: TGTTGTACATCCCTGGAAGTTG | |||||||
GAPDH | F: GTATTGTTGAGGGACTGATGACC | 182 | 57 | 0.047 | 92 | 0.999 | 23.31 |
(JN599100) | R: AGTAAGCTTGCCATTGAACTCAG | ||||||
TUB | F: TACTGTGGTTGAGCCATACAATG | 162 | 60 | 0.069 | 98 | 0.993 | 23.87 |
(JN599101) | R: GCAGAGATCAAATGGTTCAAATC | ||||||
EF1 | F: CAAGTACTACTGCACCGTCATTG | 199 | 57 | 0.025 | 89 | 0.984 | 25.10 |
(JN599095) | R: GATCATCTGCTTCACTCCAAGAG | ||||||
UBQ | F: GCAAGAAGAAGACCTACACCAAG | 225 | 60 | 0.054 | 100 | 0.991 | 19.00 |
(JN599096) | R: CCTTCTGGTTGTAGACGTAGGTG | ||||||
18S | F: GTCCAGACATAGGAAGGATTGAC | 245 | 63 | 0.052 | 106 | 0.995 | 15.08 |
(JN599097) | R: GAACATCTAAGGGCATCACAGAC | ||||||
25S | F: GCATGAATGGATTAACGAGATTC | 165 | 63 | 0.098 | 95 | 0.998 | 17.00 |
(JN599099) | R: GGCTCCCACTTATCCTACAC | ||||||
26S | F: GATAGCGTACAAGTACCGTGAGG | 238 | 63 | 0.102 | 94 | 0.993 | 20.45 |
(JN599098) | R: GTTTCGGGTCAAATAGGAAGAAC |