Selection and evaluation of optimal reference genes for quantitative reverse transcription-polymerase chain reaction analyses of gene expression in human spermatozoa
Objective: Optimal reference genes are critical for accurate normalization and reliable interpretation of gene expression quantification data. Recently, several strategies have been utilized for validating reference genes in different human tissues. However, no universal reference genes have been de...
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doaj-88e6f60baf03428182b645aabc55cec82021-02-19T09:41:01ZengWolters Kluwer Medknow PublicationsReproductive and Developmental Medicine2096-29242589-87282020-01-014421221810.4103/2096-2924.305932Selection and evaluation of optimal reference genes for quantitative reverse transcription-polymerase chain reaction analyses of gene expression in human spermatozoaChun-Hai LuoYun-Ge TangShi-Hao HongYuan TangYing ZhangFei SunObjective: Optimal reference genes are critical for accurate normalization and reliable interpretation of gene expression quantification data. Recently, several strategies have been utilized for validating reference genes in different human tissues. However, no universal reference genes have been described that accurately summarize transcriptional activity in human spermatozoa. Methods: Using quantitative reverse transcription-polymerase chain reaction (RT-qPCR), we evaluated ten commonly used candidate reference genes between two groups of human cryopreserved donor sperm with different pregnancy rates. We assessed the stability of reference genes using three different algorithms, namely geNorm, NormFinder, and BestKeeper. We then identified the most stable reference genes. Results: Male-enhanced antigen 1 (MEA1) was identified as the most stably expressed reference gene, followed by testis-enhanced gene transcript (TEGT). Conclusions: We comprehensively identified MEA1 and TEGT as the most stably expressed reference genes for the normalization of gene expression data in human spermatozoa.http://www.repdevmed.org/article.asp?issn=2096-2924;year=2020;volume=4;issue=4;spage=212;epage=218;aulast=human spermatozoa; quantitative reverse transcription-polymerase chain reaction; reference gene |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Chun-Hai Luo Yun-Ge Tang Shi-Hao Hong Yuan Tang Ying Zhang Fei Sun |
spellingShingle |
Chun-Hai Luo Yun-Ge Tang Shi-Hao Hong Yuan Tang Ying Zhang Fei Sun Selection and evaluation of optimal reference genes for quantitative reverse transcription-polymerase chain reaction analyses of gene expression in human spermatozoa Reproductive and Developmental Medicine human spermatozoa; quantitative reverse transcription-polymerase chain reaction; reference gene |
author_facet |
Chun-Hai Luo Yun-Ge Tang Shi-Hao Hong Yuan Tang Ying Zhang Fei Sun |
author_sort |
Chun-Hai Luo |
title |
Selection and evaluation of optimal reference genes for quantitative reverse transcription-polymerase chain reaction analyses of gene expression in human spermatozoa |
title_short |
Selection and evaluation of optimal reference genes for quantitative reverse transcription-polymerase chain reaction analyses of gene expression in human spermatozoa |
title_full |
Selection and evaluation of optimal reference genes for quantitative reverse transcription-polymerase chain reaction analyses of gene expression in human spermatozoa |
title_fullStr |
Selection and evaluation of optimal reference genes for quantitative reverse transcription-polymerase chain reaction analyses of gene expression in human spermatozoa |
title_full_unstemmed |
Selection and evaluation of optimal reference genes for quantitative reverse transcription-polymerase chain reaction analyses of gene expression in human spermatozoa |
title_sort |
selection and evaluation of optimal reference genes for quantitative reverse transcription-polymerase chain reaction analyses of gene expression in human spermatozoa |
publisher |
Wolters Kluwer Medknow Publications |
series |
Reproductive and Developmental Medicine |
issn |
2096-2924 2589-8728 |
publishDate |
2020-01-01 |
description |
Objective: Optimal reference genes are critical for accurate normalization and reliable interpretation of gene expression quantification data. Recently, several strategies have been utilized for validating reference genes in different human tissues. However, no universal reference genes have been described that accurately summarize transcriptional activity in human spermatozoa.
Methods: Using quantitative reverse transcription-polymerase chain reaction (RT-qPCR), we evaluated ten commonly used candidate reference genes between two groups of human cryopreserved donor sperm with different pregnancy rates. We assessed the stability of reference genes using three different algorithms, namely geNorm, NormFinder, and BestKeeper. We then identified the most stable reference genes.
Results: Male-enhanced antigen 1 (MEA1) was identified as the most stably expressed reference gene, followed by testis-enhanced gene transcript (TEGT).
Conclusions: We comprehensively identified MEA1 and TEGT as the most stably expressed reference genes for the normalization of gene expression data in human spermatozoa. |
topic |
human spermatozoa; quantitative reverse transcription-polymerase chain reaction; reference gene |
url |
http://www.repdevmed.org/article.asp?issn=2096-2924;year=2020;volume=4;issue=4;spage=212;epage=218;aulast= |
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