scholarly journals Selection of reference genes suitable for normalization of RT-qPCR data in glioma stem cells

BioTechniques ◽  
2020 ◽  
Vol 68 (3) ◽  
pp. 130-137
Author(s):  
Weiqi Dang ◽  
Xiang Zhang ◽  
Qinghua Ma ◽  
Lu Chen ◽  
Mianfu Cao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Reference Genes ◽  
Geometric Mean ◽  
Specific Reference ◽  
Glioma Stem Cells ◽  
Reverse Transcription Pcr ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Suitable Reference

Considering the importance of gene expression studies for understanding the biology of glioma stem cells (GSCs), we aimed to identify the reliable reference genes in GSCs that were derived from the glioma cell lines T98G, LN229, 090116 and 091214. Quantitative real-time reverse-transcription PCR was employed using 11 reference genes identified through a PubMed literature search, and the assessment of stability through the geNorm, Normfinder and coefficient of variation methods was performed to select suitable reference genes. We found that HPRT1 and RPL13A were the most suitable reference genes, and validated the geometric mean of these genes to normalize the expression of stemness genes by GSCs. Therefore, it is necessary to select novel cell-specific reference genes with greater expression stability for gene expression studies in GSCs.

scholarly journals Selection and Validation of Appropriate Reference Genes for Gene Expression Studies in Schima Superba

2021 ◽  
Author(s):  
Zhongyi Yang ◽  
Rui Zhang ◽  
Zhichun Zhou
Keyword(s):  
Gene Expression ◽  
Reference Gene ◽  
Reference Genes ◽  
Geometric Mean ◽  
Gene Transcript ◽  
Expression Stability ◽  
Schima Superba ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Different Tissues

Abstract Background Quantitative real-time PCR (qRT-PCR) is a reliable and high-throughput technique for gene expression studies, but its accuracy depends on the expression stability of reference genes. Schima superba is a strong resistance and fast-growing timber specie. However, so far, reliable reference gene identifications have not been reported in S. superba. In this study, we screened and verified the stably expressed reference genes in different tissues of S. superba.Results Nineteen candidate reference genes were selected and evaluated for their expression stability in different tissues. Three software programs (geNorm, NormFinder, and BestKeeper) were used to evaluate the reference gene transcript stabilities, and comprehensive stability ranking was generated by the geometric mean method. Our results identified that SsuACT was the most stable reference gene, SsuACT + SsuRIB was the best reference genes combination for different tissues. Finally, the stable and less stable reference genes were verified using the SsuSND1 expression in different tissues.Conclusions This is the first report to verify the appropriate reference genes for normalizing gene expression in S. superba for different tissues, which will facilitate future elucidation of gene regulations in this species, and useful references for relative species.

scholarly journals Identification of Suitable Reference Genes for gene Expression Studies by qRT-PCR in the Blister BeetleMylabris cichorii

2014 ◽  
Vol 14 (94) ◽  
pp. 1-10 ◽  
Author(s):  
Yu Wang ◽  
Zhong-Kang Wang ◽  
Yi Huang ◽  
Yu-Feng Liao ◽  
You-Ping Yin
Keyword(s):  
Gene Expression ◽  
Reference Genes ◽  
Qrt Pcr ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Suitable Reference

scholarly journals In search of suitable reference genes for gene expression studies of human renal cell carcinoma by real-time PCR

2007 ◽  
Vol 8 (1) ◽  
pp. 47 ◽  
Author(s):  
Monika Jung ◽  
Azizbek Ramankulov ◽  
Jan Roigas ◽  
Manfred Johannsen ◽  
Martin Ringsdorf ◽  
...  
Keyword(s):  
Gene Expression ◽  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Real Time Pcr ◽  
Renal Cell ◽  
Reference Genes ◽  
Human Renal Cell Carcinoma ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Suitable Reference

Determination of Suitable Candidate Genes as Endogenous Control for Gene Expression Analysis in Local Capsicum Annuum MC11

2021 ◽  
Vol 12 (3) ◽  
pp. 1011-1017
Author(s):  
Marina Mokhtar Et.al
Keyword(s):  
Gene Expression ◽  
Candidate Genes ◽  
Capsicum Annuum ◽  
Reference Genes ◽  
Data Normalization ◽  
Plant Parts ◽  
Qrt Pcr ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Suitable Reference

Quantitative real-time polymerase chain reaction (qRT-PCR) is one of the most common methods for gene expression studies. Data normalization based on reference genes is essential for qRT-PCR assays. This study identifies suitable reference genes for local chilli, Capsicum annuum var MC11 under incident of Cucumber mosaic virus infection. Six candidate genes actin, tub, EF1α, GAPDH, TEF1α and 18SrRNA and three validated Capsicum reference genes UBI-3 ref, β-tub ref and gapdhref were tested against five chilli plant parts stem, shoot, leave, flower and root.  The PCR/qRT-PCR results demonstrate only five candidate references genes actin, EF1α, GAPDH, 18SrRNA, and TEF1α that show specific single band of amplicon, without primer dimers and at the targeted sizes. Through qRT-PCR, GAPDH gives single peak in dissociation curve in all plant parts used further fulfilling the characteristic of reference genes.Previous work on validation of reference genes in pepper shows that only UBI-3 suits to C. annuum var MC11 infected CMV, thus we suggest that GAPDH has a potential to be a validated reference gene for C. annuum var MC11 and can be used together UBI-3 for the purpose of data normalization. 

Identification of Suitable Reference Genes for Gene Expression Studies in Tissues from Fructose-Fed Rats

2012 ◽  
Vol 5 (2) ◽  
pp. 560-565 ◽  
Author(s):  
Ana Djordjevic ◽  
Danijela Vojnović Milutinović ◽  
Nikola Tanić ◽  
Biljana Bursić ◽  
Ana Vasiljević ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reference Genes ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Suitable Reference

scholarly journals Evaluation of suitable reference genes for gene expression studies in porcine alveolar macrophages in response to LPS and LTA

2012 ◽  
Vol 5 (1) ◽  
pp. 107 ◽  
Author(s):  
Mehmet Ulas Cinar ◽  
Mohammad Ariful Islam ◽  
Muhammad Jasim Uddin ◽  
Ernst Tholen ◽  
Dawit Tesfaye ◽  
...  
Keyword(s):  
Gene Expression ◽  
Alveolar Macrophages ◽  
Reference Genes ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Suitable Reference
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