Wheat-Specific Gene,Ribosomal Protein L21, Used as the Endogenous Reference Gene for Qualitative and Real-Time Quantitative Polymerase Chain Reaction Detection of Transgenes

2014 ◽  
Vol 62 (43) ◽  
pp. 10405-10413 ◽  
Author(s):  
Yi-Ke Liu ◽  
He-Ping Li ◽  
Tao Huang ◽  
Wei Cheng ◽  
Chun-Sheng Gao ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Ribosomal Protein ◽  
Reference Gene ◽  
Quantitative Polymerase Chain Reaction ◽  
Specific Gene ◽  
Polymerase Chain Reaction Detection ◽  
Chain Reaction ◽  
Endogenous Reference Gene ◽  
Endogenous Reference ◽  
Polymerase Chain

scholarly journals Validation of an appropriate reference gene for normalization of reverse transcription–quantitative polymerase chain reaction data from rectal cancer biopsies

2009 ◽  
Vol 388 (2) ◽  
pp. 348-350 ◽  
Author(s):  
Alexandre Ho-Pun-Cheung ◽  
Caroline Bascoul-Mollevi ◽  
Eric Assenat ◽  
Frédéric Bibeau ◽  
Florence Boissière-Michot ◽  
...  
Keyword(s):  
Rectal Cancer ◽  
Polymerase Chain Reaction ◽  
Reference Gene ◽  
Reverse Transcription ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Reaction Data ◽  
Polymerase Chain ◽  
Polymerase Chain Reaction Data

scholarly journals TATA box binding protein and ribosomal protein 4 are suitable reference genes for normalization during quantitative polymerase chain reaction study in bovine mesenchymal stem cells

2020 ◽  
Vol 33 (12) ◽  
pp. 2021-2030
Author(s):  
Si-Jung Jang ◽  
Ryoung-Hoon Jeon ◽  
Hwan-Deuk Kim ◽  
Jong-Chan Hwang ◽  
Hyeon-Jeong Lee ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Ribosomal Protein ◽  
Reference Genes ◽  
Binding Protein ◽  
Quantitative Polymerase Chain Reaction ◽  
Tata Box ◽  
Chain Reaction ◽  
Experimental Conditions ◽  
Polymerase Chain ◽  
Tata Box Binding Protein

Objective: Quantitative polymerase chain reaction (qPCR) has been extensively used in the field of mesenchymal stem cell (MSC) research to elucidate their characteristics and clinical potential by normalization of target genes against reference genes (RGs), which are believed to be stably expressed irrespective of various experimental conditions. However, the expression of RGs is also variable depending on the experimental conditions, which may lead to false or contradictory conclusions upon normalization. Due to the current lack of information for a clear list of stable RGs in bovine MSCs, we conducted this study to identify suitable RGs in bovine MSCs.Methods: The cycle threshold values of ten traditionally used RGs (18S ribosomal RNA [18S], beta-2-microglobulin [B2M], H2A histone family, member Z [H2A], peptidylprolyl isomerase A [PPIA], ribosomal protein 4 [RPL4], succinate dehydrogenase complex, subunit A [SDHA], beta actin [ACTB], glyceraldehyde-3-phosphate dehydrogenase [GAPDH], TATA box binding protein [TBP], and hypoxanthine phosphoribosyltrasnfrase1 [HPRT1]) in bovine bone marrow-derived MSCs (bBMMSCs) were validated for their stabilities using three types of RG evaluation algorithms (geNorm, Normfinder, and Bestkeeper). The effect of validated RGs was then verified by normalization of lineage-specific genes (fatty acid binding protein 4 [FABP4] and osteonectin [ON]) expressions during differentiations of bBMMSCs or POU class 5 homeobox 1 (OCT4) expression between bBMMSCs and dermal skins.Results: Based on the results obtained for the three most stable RGs from geNorm (TBP, RPL4, and H2A), Normfinder (TBP, RPL4, and SDHA), and Bestkeeper (TBP, RPL4, and SDHA), it was comprehensively determined that TBP and RPL4 were the most stable RGs in bBMMSCs. However, traditional RGs were suggested to be the least stable (18S) or moderately stable (GAPDH and ACTB) in bBMMSCs. Normalization of FABP4 or ON against TBP, RPL4, and 18S presented significant differences during differentiation of bBMMSCs. However, although significantly low expression of OCT4 was detected in dermal skins compared to that in bBMMSCs when TBP and RPL4 were used in normalization, normalization against 18S exhibited no significance.Conclusion: This study proposes that TBP and RPL4 were suitable as stable RGs for qPCR study in bovine MSCs.

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