Table 1.
Eight candidate reference genes assessed for gene expression normalisation in Olea europaea and amplicon characteristics
| Gene name | Accession number/cluster ID* | Primer sequence (5' to 3') | Amplicon length (bp) | Annealing temperature** (°C) | PCR efficiency value*** | Standard error (SE) | R2 **** |
|---|---|---|---|---|---|---|---|
|
60S RBP L18-3 |
[Oleadatabase:Cluster ID-OLEEUCl011221] contig 2 |
F: GTAAGAGCAAGAAGACCAAG |
101 |
55 |
1.984 |
0.014 |
0.978 |
| R: GCTTCCAGTTCTCCTCAC | |||||||
|
PP2A |
[Oleadatabase:Cluster ID-OLEEUCl010038] contig 1 |
F: AGATCGGTGAAATACTTCCACACG |
189 |
56 |
2.038 |
0.020 |
0.969 |
| R: TCGTGGATACTACTCAGTGGAGACTG | |||||||
|
PTB |
[Oleadatabase:Cluster ID-OLEEUCl031691] contig 1 |
F: CTTCTCCGAAATAAACCAGAT |
156 |
56 |
2.243 |
0.057 |
0.855 |
| R: GGTGTCAGCTCCAGTTGTAA | |||||||
|
TUBA |
[Oleadatabase:Cluster ID-OLEEUCl051890] contig 1 |
F: AGAACACCTCAGCAACAC |
100 |
51 |
1.704 |
0.069 |
0.870 |
| R: AACTACCAGCCACCAACT | |||||||
|
TIP2 |
[Oleadatabase:Cluster ID-OLEEUCl011159] contig 2 |
F: ACTTGTTGTAAGCAATGG |
104 |
51 |
2.093 |
0.081 |
0.935 |
| R: TGATTCATTAAGCGTTGG | |||||||
|
OUB2 |
[GenBank:AF429430.1] |
F: AATGAAGTCTGTGTGTCCTTTGG |
150 |
51 |
1.513 |
0.054 |
0.890 |
| R: AAGGGAAATCCCATCAACG | |||||||
|
GAPDH |
[GenBank:EF506530.1] |
F: ACAGCTCCTGGTAAGGGTGA |
210 |
56 |
2.111 |
0.018 |
0.971 |
| R: GGCTTGCGTCAAGAAGTCTC | |||||||
| EF1-alpha | [GenBank:XM_002527974.1] | F: GAATGGTGATGCTGGTTTCG |
191 | 56 | 1.908 | 0.005 | 0.996 |
| R: CCCTTCTTGGCAGCAGACTTG |
*Cluster ID obtained from the Olea database [26].
**Annealing temperature used in PCR experiments.
***Measure of the PCR amplification efficiency calculated from calibration curve derived from prepared standards.
****correlation coefficient (R2) for the calibration curve.