UBQUniversitat de Barcelona-Químiques (Ciencias)
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The six candidate genes selected for normalization ([alpha]TUB, GAPDH, H1, SAMDC, UBQ, and 25S rRNA) in RT-qPCR tests showed Cq values ranging from 13.06 to 28.00 (Supplemental Table S1).
The two software programs identified histone H1 (M = 0.28; CV [+ or -] SD = 1.06 [+ or -] 0.26) and [alpha]TUB (M = 0.32; CV [+ or -] SD = 1.06 [+ or -] 0.29) as the most stable genes, followed by GAPDH, 25S rRNA, UBQ, and SAMDC (see respective values in Table 3).
In the present study, a group of potential reference genes ([alpha]TUB, GAPDH, H1, SAMDC, UBQ, and 25S rRNA) were evaluated by the three software programs, in order to evaluate their reliability for expression profiles normalization in sugarcane roots under abiotic stress (24 h of continuous dehydration).
ACT, EF1, and UBQ genes were selected as internal controls to normalize the level of total RNA present in each reaction [17].