Validation of reference genes in Solenopsis invicta in different developmental stages, castes and tissues.
To accurately assess gene expression levels, it is essential to normalize real-time quantitative PCR (RT-qPCR) data with suitable internal reference genes. For the red imported fire ant, Solenopsis invicta, reliable reference genes to assess the transcript expression levels of the target genes have...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3585193?pdf=render |
| _version_ | 1819089583147581440 |
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| author | Daifeng Cheng Zhiling Zhang Xiaofang He Guangwen Liang |
| author_facet | Daifeng Cheng Zhiling Zhang Xiaofang He Guangwen Liang |
| author_sort | Daifeng Cheng |
| collection | DOAJ |
| description | To accurately assess gene expression levels, it is essential to normalize real-time quantitative PCR (RT-qPCR) data with suitable internal reference genes. For the red imported fire ant, Solenopsis invicta, reliable reference genes to assess the transcript expression levels of the target genes have not been previously investigated. In this study, we examined the expression levels of five candidate reference genes (rpl18, ef1-beta, act, GAPDH, and tbp) in different developmental stages, castes and tissues of S. invicta. To evaluate the suitability of these genes as endogenous controls, three software-based approaches (geNorm, BestKeeper and NormFinder) and one web-based comprehensive tool (RefFinder) were used to analyze and rank the tested genes. Furthermore, the optimal number of reference gene(s) was determined by the pairwise variation value. Our data showed that two of the five candidate genes, rpl18 and ef1-beta, were the most suitable reference genes because they have the most stable expression among different developmental stages, castes and tissues in S. invicta. Although widely used as reference gene in other species, in S. invicta the act gene has high variation in expression and was consequently excluded as a reliable reference gene. The two validated reference genes, rpl18 and ef1-beta, can be widely used for quantification of target gene expression with RT-qPCR technology in S. invicta. |
| first_indexed | 2024-12-21T22:10:14Z |
| format | Article |
| id | doaj.art-11f9e8cbf2404cbcbd420b287cdd7cb4 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-21T22:10:14Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-11f9e8cbf2404cbcbd420b287cdd7cb42022-12-21T18:48:36ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0182e5771810.1371/journal.pone.0057718Validation of reference genes in Solenopsis invicta in different developmental stages, castes and tissues.Daifeng ChengZhiling ZhangXiaofang HeGuangwen LiangTo accurately assess gene expression levels, it is essential to normalize real-time quantitative PCR (RT-qPCR) data with suitable internal reference genes. For the red imported fire ant, Solenopsis invicta, reliable reference genes to assess the transcript expression levels of the target genes have not been previously investigated. In this study, we examined the expression levels of five candidate reference genes (rpl18, ef1-beta, act, GAPDH, and tbp) in different developmental stages, castes and tissues of S. invicta. To evaluate the suitability of these genes as endogenous controls, three software-based approaches (geNorm, BestKeeper and NormFinder) and one web-based comprehensive tool (RefFinder) were used to analyze and rank the tested genes. Furthermore, the optimal number of reference gene(s) was determined by the pairwise variation value. Our data showed that two of the five candidate genes, rpl18 and ef1-beta, were the most suitable reference genes because they have the most stable expression among different developmental stages, castes and tissues in S. invicta. Although widely used as reference gene in other species, in S. invicta the act gene has high variation in expression and was consequently excluded as a reliable reference gene. The two validated reference genes, rpl18 and ef1-beta, can be widely used for quantification of target gene expression with RT-qPCR technology in S. invicta.http://europepmc.org/articles/PMC3585193?pdf=render |
| spellingShingle | Daifeng Cheng Zhiling Zhang Xiaofang He Guangwen Liang Validation of reference genes in Solenopsis invicta in different developmental stages, castes and tissues. PLoS ONE |
| title | Validation of reference genes in Solenopsis invicta in different developmental stages, castes and tissues. |
| title_full | Validation of reference genes in Solenopsis invicta in different developmental stages, castes and tissues. |
| title_fullStr | Validation of reference genes in Solenopsis invicta in different developmental stages, castes and tissues. |
| title_full_unstemmed | Validation of reference genes in Solenopsis invicta in different developmental stages, castes and tissues. |
| title_short | Validation of reference genes in Solenopsis invicta in different developmental stages, castes and tissues. |
| title_sort | validation of reference genes in solenopsis invicta in different developmental stages castes and tissues |
| url | http://europepmc.org/articles/PMC3585193?pdf=render |
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