Selection of internal references for RT-qPCR assays in Neurofibromatosis type 1 (NF1) related Schwann cell lines.
Real-time quantitative PCR (RT-qPCR) has been widely applied in uncovering disease mechanisms and screening potential biomarkers. Internal reference gene selection determines the accuracy and reproducibility of data analyses. The aim of this study was to identify the optimal reference genes for the...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2021-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0241821 |
| _version_ | 1818690698619125760 |
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| author | Yi-Hui Gu Xi-Wei Cui Jie-Yi Ren Man-Mei Long Wei Wang Cheng-Jiang Wei Rehanguli Aimaier Yue-Hua Li Man-Hon Chung Bin Gu Qing-Feng Li Zhi-Chao Wang |
| author_facet | Yi-Hui Gu Xi-Wei Cui Jie-Yi Ren Man-Mei Long Wei Wang Cheng-Jiang Wei Rehanguli Aimaier Yue-Hua Li Man-Hon Chung Bin Gu Qing-Feng Li Zhi-Chao Wang |
| author_sort | Yi-Hui Gu |
| collection | DOAJ |
| description | Real-time quantitative PCR (RT-qPCR) has been widely applied in uncovering disease mechanisms and screening potential biomarkers. Internal reference gene selection determines the accuracy and reproducibility of data analyses. The aim of this study was to identify the optimal reference genes for the relative quantitative analysis of RT-qPCR in fourteen NF1 related cell lines, including non-tumor, benign and malignant Schwann cell lines. The expression characteristics of eleven candidate reference genes (RPS18, ACTB, B2M, GAPDH, PPIA, HPRT1, TBP, UBC, RPLP0, TFRC and RPL32) were screened and analyzed by four software programs: geNorm, NormFinder, BestKeeper and RefFinder. Results showed that GAPDH, the most frequently used internal reference gene, was significantly unstable between various cell lines. The combinational use of two reference genes (PPIA and TBP) was optimal in malignant Schwann cell lines and the use of single reference genes (PPIA or PRLP0) alone or in combination was optimal in benign Schwann cell lines. These recommended internal reference gene selections may improve the accuracy and reproducibility of RT-qPCR in gene expression analyses of NF1 related tumors. |
| first_indexed | 2024-12-17T12:30:08Z |
| format | Article |
| id | doaj.art-0d46e7e3a6b74bc39e5cd1badff99d64 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-17T12:30:08Z |
| publishDate | 2021-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-0d46e7e3a6b74bc39e5cd1badff99d642022-12-21T21:48:37ZengPublic Library of Science (PLoS)PLoS ONE1932-62032021-01-01162e024182110.1371/journal.pone.0241821Selection of internal references for RT-qPCR assays in Neurofibromatosis type 1 (NF1) related Schwann cell lines.Yi-Hui GuXi-Wei CuiJie-Yi RenMan-Mei LongWei WangCheng-Jiang WeiRehanguli AimaierYue-Hua LiMan-Hon ChungBin GuQing-Feng LiZhi-Chao WangReal-time quantitative PCR (RT-qPCR) has been widely applied in uncovering disease mechanisms and screening potential biomarkers. Internal reference gene selection determines the accuracy and reproducibility of data analyses. The aim of this study was to identify the optimal reference genes for the relative quantitative analysis of RT-qPCR in fourteen NF1 related cell lines, including non-tumor, benign and malignant Schwann cell lines. The expression characteristics of eleven candidate reference genes (RPS18, ACTB, B2M, GAPDH, PPIA, HPRT1, TBP, UBC, RPLP0, TFRC and RPL32) were screened and analyzed by four software programs: geNorm, NormFinder, BestKeeper and RefFinder. Results showed that GAPDH, the most frequently used internal reference gene, was significantly unstable between various cell lines. The combinational use of two reference genes (PPIA and TBP) was optimal in malignant Schwann cell lines and the use of single reference genes (PPIA or PRLP0) alone or in combination was optimal in benign Schwann cell lines. These recommended internal reference gene selections may improve the accuracy and reproducibility of RT-qPCR in gene expression analyses of NF1 related tumors.https://doi.org/10.1371/journal.pone.0241821 |
| spellingShingle | Yi-Hui Gu Xi-Wei Cui Jie-Yi Ren Man-Mei Long Wei Wang Cheng-Jiang Wei Rehanguli Aimaier Yue-Hua Li Man-Hon Chung Bin Gu Qing-Feng Li Zhi-Chao Wang Selection of internal references for RT-qPCR assays in Neurofibromatosis type 1 (NF1) related Schwann cell lines. PLoS ONE |
| title | Selection of internal references for RT-qPCR assays in Neurofibromatosis type 1 (NF1) related Schwann cell lines. |
| title_full | Selection of internal references for RT-qPCR assays in Neurofibromatosis type 1 (NF1) related Schwann cell lines. |
| title_fullStr | Selection of internal references for RT-qPCR assays in Neurofibromatosis type 1 (NF1) related Schwann cell lines. |
| title_full_unstemmed | Selection of internal references for RT-qPCR assays in Neurofibromatosis type 1 (NF1) related Schwann cell lines. |
| title_short | Selection of internal references for RT-qPCR assays in Neurofibromatosis type 1 (NF1) related Schwann cell lines. |
| title_sort | selection of internal references for rt qpcr assays in neurofibromatosis type 1 nf1 related schwann cell lines |
| url | https://doi.org/10.1371/journal.pone.0241821 |
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