Selection of reliable reference genes for gene expression studies in peach using real-time PCR
作者： Zhaoguo Tong , Zhihong Gao , Fei Wang , Jun Zhou , Zhen Zhang
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摘要: RT-qPCR is a preferred method for rapid and reliable quantification of gene expression studies. Appropriate application in such studies requires the use reference gene(s) as an internal control to normalize mRNA levels between different samples exact comparison level. However, recent have shown that no single universal all experiments. Thus, identification high quality paramount importance interpretation data generated by RT-qPCR. Only few on genes been done plants none peach (Prunus persica L. Batsch). Therefore, present study was conducted identify suitable normalization peach. In this work, eleven were investigated using with SYBR green. These are: actin 2/7 (ACT), cyclophilin (CYP2), RNA polymerase II (RP II), phospholipase A2 (PLA2), ribosomal protein L13 (RPL13), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 18S (18S rRNA), tubblin beta (TUB), alpha (TUA), translation elongation factor 2 (TEF2) ubiquitin 10 (UBQ10). All displayed wide range Cq values samples, indicating they expressed variably. The stability these except RPL13 determined three descriptive statistics, geNorm, NormFinder BestKeeper, which produced highly comparable results. Our demonstrates varied greatly studied Based results from BestKeeper analyses, sample pools analyzed, TEF2, UBQ10 RP found be most very statistical reliability, TEF2 other series, while rRNA, PLA2 unsuitable controls. GAPDH ACT also performed poorly less stable our analysis. To achieve accurate expression, two or more must used normalization. combinations TEF2/UBQ10/RP TEF2/RP suggested subsets, respectively.
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