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Evaluation of putative internal reference genes for gene expression normalization in Nannochloropsis sp by quantitative real-time RT-PCR
Cao S1,2; Zhang X1; Ye N1; Fan X1; Mou S1; Xu D1; Liang C3; Wang Y2; Wang W2
2012
Source PublicationBIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
ISSN0006-291X
Volume424Issue:1Pages:118-123
AbstractQuantitative real-time reverse transcription PCR (RT-qPCR), a sensitive technique for quantifying gene expression, depends on the stability of the reference gene(s) used for data normalization. To date, few studies on reference genes have been undertaken for Nannochloropsis sp. In this study, 12 potential reference genes were evaluated for their expression stability using the geNorm and NormFinder statistical algorithms by RT-qPCR. The results showed that the best reference genes differed depending on the treatments: different light intensities (DL), the diurnal cycle (DC), high light intensity (HL) and low temperature treatments (LT). A combination of ACT1, ACT2 and TUA would be appropriate as a reference panel for normalizing gene expression data across all the treatments. ACT2 showed the most stable expression across all tested samples but was not the most stable one for individual treatments. Though 18S showed the least stable expression considering all tested samples, it is the most stable one for LT using geNorm. The expression of Lhc confirmed that the appropriate reference genes are crucial. These results provide a foundation for more accurate use of RT-qPCR under different experimental conditions in Nannochloropsis sp. gene analysis.
KeywordReference Genes Nannochloropsis Sp Gene Expression Normalization Rt-qpcr
DOI10.1016/j.bbrc.2012.06.086
Indexed BySCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology ; Biophysics
WOS SubjectBiochemistry & Molecular Biology ; Biophysics
WOS IDWOS:000307135700021
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Cited Times [WOS]:27   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionInstitute of Chinese Medical Sciences
Corresponding AuthorYe N
Affiliation1.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China.
2.Qingdao Agr Univ, Qingdao 266109, Peoples R China
3.Qingdao Univ Sci & Technol, Qingdao 266042, Peoples R China
Recommended Citation
GB/T 7714
Cao S,Zhang X,Ye N,et al. Evaluation of putative internal reference genes for gene expression normalization in Nannochloropsis sp by quantitative real-time RT-PCR[J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,2012,424(1):118-123.
APA Cao S.,Zhang X.,Ye N.,Fan X.,Mou S.,...&Wang W.(2012).Evaluation of putative internal reference genes for gene expression normalization in Nannochloropsis sp by quantitative real-time RT-PCR.BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,424(1),118-123.
MLA Cao S,et al."Evaluation of putative internal reference genes for gene expression normalization in Nannochloropsis sp by quantitative real-time RT-PCR".BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 424.1(2012):118-123.
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