Table 6.
Comprehensive ranking and best recommended pair of reference genes for accurate normalization of qPCR assays using samples of Meliponini bees under different experimental contexts. The comprehensive ranking is based on gene expression stability of each gene calculated by RefFinder using geNorm, NormFinder, BestKeeper and delta-Ct algorithms.
| Condition | Reference Gene | F. varia | M. quadrifasciata | S. bipunctata | ||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Comprehensive Ranking | Most stable | Comprehensive Ranking | Most stable | Comprehensive Ranking | Most stable | |||||
| Stability | Rank | Stability | Rank | Stability | Rank | |||||
| Development | ef1-α | 5.23 | 6 | rpl32, rps18 | 5.73 | 6 | rpl32, rps18 | 5.23 | 6 | rpl32, rps18 |
| gapdh | 4.56 | 5 | 5.23 | 5 | 3.83 | 4 | ||||
| rpl32 | 1.41 | 1 | 1.57 | 1 | 1.41 | 1 | ||||
| rps5 | 2.83 | 3 | 2.21 | 3 | 3.22 | 3 | ||||
| rps18 | 2.06 | 2 | 2.06 | 2 | 1.57 | 2 | ||||
| tbp-af | 3.08 | 4 | 2.83 | 4 | 4.23 | 5 | ||||
| Sex | ef1-α | 6.00 | 6 | rps5, tbp-af | 6.00 | 6 | rpl32, rps18 | 6.00 | 6 | rps18, rpl32 |
| gapdh | 3.36 | 5 | 5.00 | 5 | 4.00 | 4 | ||||
| rpl32 | 3.22 | 3 | 1.32 | 1 | 1.57 | 2 | ||||
| rps5 | 1.50 | 1 | 2.28 | 3 | 4.40 | 5 | ||||
| rps18 | 3.34 | 4 | 1.68 | 2 | 1.19 | 1 | ||||
| tbp-af | 1.86 | 2 | 4.00 | 4 | 3.08 | 3 | ||||
| Tissues, organs and body parts | ef1-α | 5.73 | 6 | rps18, rps5 | 4.47 | 5 | rps18, tbp-af | 6.00 | 6 | rps5, tbp-af |
| gapdh | 5.23 | 5 | 6.00 | 6 | 5.00 | 5 | ||||
| rpl32 | 2.63 | 3 | 2.06 | 3 | 2.06 | 3 | ||||
| rps5 | 2.06 | 2 | 3.16 | 4 | 2.00 | 1 | ||||
| rps18 | 1.41 | 1 | 1.86 | 1 | 2.45 | 4 | ||||
| tbp-af | 2.63 | 4 | 1.86 | 2 | 2.00 | 2 | ||||
| Bacterial injection | ef1-α | 6.00 | 6 | rps18, rpl32 | 6.00 | 6 | rps18, tbp-af | 6.00 | 6 | rps18, rpl32 |
| gapdh | 5.00 | 5 | 5.00 | 5 | 4.23 | 4 | ||||
| rpl32 | 1.86 | 2 | 3.13 | 4 | 1.41 | 2 | ||||
| rps5 | 2.63 | 3 | 2.45 | 3 | 4.40 | 5 | ||||
| rps18 | 1.57 | 1 | 1.57 | 1 | 1.19 | 1 | ||||
| tbp-af | 2.63 | 4 | 1.68 | 2 | 3.22 | 3 | ||||
| Pesticide exposure | ef1-α | 5.00 | 5 | tbp-af, rps5 | 4.40 | 5 | rpl32, rps18 | 3.98 | 5 | rps5, tbp-af |
| gapdh | 6.00 | 6 | 6.00 | 6 | 6.00 | 6 | ||||
| rpl32 | 3.13 | 3 | 1.68 | 1 | 3.94 | 4 | ||||
| rps5 | 1.57 | 2 | 3.66 | 4 | 1.41 | 1 | ||||
| rps18 | 3.13 | 4 | 1.86 | 2 | 2.21 | 3 | ||||
| tbp-af | 1.32 | 1 | 2.00 | 3 | 2.06 | 2 | ||||