Gene (coffee | Reference | Primer sequence | Amplicon length (bp) | Ta (°C) | Tm (°C) | PCR efficiency (%) | Regression coefficient (R2) | Average Cq value |
---|---|---|---|---|---|---|---|---|
source gene) | ||||||||
VATP16 (GT00156.1)* | Gamm et al. [29] | Fw: ATAGAGAGAGAGCCCCCAATTC | 150 | 60 | 78.63 | Discarded (unspecific amplification, primer dimer) | ||
Rev: CCGAGGAAGCCAAAAAAAG | ||||||||
UQCC (isotig09120) | Fw: CCTCGGGCTTCATCTCTACTC | 156 | 60 | 77.21 | ||||
Rev: TCTCCCCAGCATTTTTTGTC | ||||||||
SAND (isotig05620) | Selim et al. [30] | Fw: GATTTGTCTACGAACCCTGCTT | 116 | 60 | 78.70 | |||
Rev: GGACCACCAACACCAATAAAC | ||||||||
UBQ9 (AF29708)* | Ramiro et al. [7] | Fw: AACATTGAGGGTGGTTCTGTTC | 79 | 60 | 76.16 | 92 | 0.987 | 24.5 ± 0.95 |
Rev: GCAGAAAACCAACTAAGACCTAACAA | ||||||||
S24 (SNG-U349723)a | Cruz et al. [28] | Fw: GCCCAAATATCGGCTTATCA | 92 | 60 | 76.60 | 91 | 0.996 | 20.61 ± 1.04 |
Rev: TCTTCTTGGCCCTGTTCTTC | ||||||||
IDE (isotig10635) | Borges et al. [12] | Fw: TGATCTAAGCTGGTGGAAAGC | 91 | 55 | 76.28 | 92 | 0.994 | 24.46 ± 1.30 |
Rev: TCAGGTGCATCAGGATGATT | ||||||||
β- Tub9 (isotig08544) | Fw: ACCCTCCAGCAAACTGATGA | 100 | 55 | 77.27 | 96 | 0.996 | 19.40 ± 0.82 | |
Rev: AGGATGCCACTGCTGATGAT | ||||||||
14-3-3 (SGN-U356404)a | Barsalobres-Cavallari et al. [18] | Fw: TGTGCTCTTTAGCTTCCAAACG | 75 | 60 | 73.33 | 103 | 0.999 | 22.76 ± 1.47 |
Rev: CTTCACGAGACATATTGTCTTACTCAAA | ||||||||
GADPH (SGN-U347734)a | Fw: TTGAAGGGCGGTGCAAA | 59 | 60 | 75.73 | 96 | 0.998 | 21.42 ± 1.96 | |
Rev: AACATGGGTGCATCCTTGCT | ||||||||
RPL7 (SGN-U351477)a | Barsalobres-Cavallari et al. [18] | Fw: CATTCGAGGTATCAATGCTATGCA | 66 | 60 | 76.48 | 89 | 0.999 | 23.76 ± 1.28 |
Rev: TGTCTCAGGCGCAGAAGCT | ||||||||
Genes of interest (GOIs) | ||||||||
PR10 (CF589103)* | Ramiro et al. [7] | Fw: GCCACCATCCTTGAAGAGAA | 151 | 55 | 80.17 | 99 | 0.999 | 19.35 ± 2.28 |
Rev: CAACTCTCTGCTTGGCAGTCT | ||||||||
RLK (CF589181)* | Fw: ATGGGAGAAAAGAATGGCAGAAG | 189 | 55 | 81.15 | 91 | 0.998 | 24.81 ± 2.02 | |
Rev: GGCCAATTACAGTTTGAAAACACC |