In:
PLOS ONE, Public Library of Science (PLoS), Vol. 16, No. 2 ( 2021-2-25), p. e0241821-
Abstract:
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.
Type of Medium:
Online Resource
ISSN:
1932-6203
DOI:
10.1371/journal.pone.0241821
DOI:
10.1371/journal.pone.0241821.g001
DOI:
10.1371/journal.pone.0241821.g002
DOI:
10.1371/journal.pone.0241821.g003
DOI:
10.1371/journal.pone.0241821.g004
DOI:
10.1371/journal.pone.0241821.g005
DOI:
10.1371/journal.pone.0241821.g006
DOI:
10.1371/journal.pone.0241821.s001
DOI:
10.1371/journal.pone.0241821.s002
DOI:
10.1371/journal.pone.0241821.s003
DOI:
10.1371/journal.pone.0241821.s004
DOI:
10.1371/journal.pone.0241821.s005
DOI:
10.1371/journal.pone.0241821.s006
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2021
detail.hit.zdb_id:
2267670-3
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