We tested the amplification efficiency before using housekeeping genes. Referring to the previous study of Arabidopsis housekeeping genes (Dheda, 2004; Czechowski, 2005; Hou, 2018), we selected Atactin1 (F: AGGCACCTCTTAACCCTAAAGC, R: GGACAACGGAATCTCTCAGC), Atactin2 (F: TTGTGCTGGATTCTGGTGATGG, R: CCGCTCTCTGCTGTTGTGGTG) and Atactin8 (F: GAATTACCCGACGGACA, R: ACGGTCTGCAATACCT) were tested for efficiency, and we found that the best result was Atactin2. The best housekeeping gene for O. fragrans is OfACT, which has been reported in a previous study (Zhang, 2015).
Auhtors report that Actin was used as housekeeping gene. Do the authors have performed an efficiency test before using it? Do they tried to use otherHK genes?