Loss of Gene Information: Discrepancies between RNA Sequencing, cDNA Microarray, and qRT-PCR.
Int J Mol Sci
; 22(17)2021 Aug 28.
Article
em En
| MEDLINE
| ID: mdl-34502254
Molecular analyses of normal and diseased cells give insight into changes in gene expression and help in understanding the background of pathophysiological processes. Years after cDNA microarrays were established in research, RNA sequencing (RNA-seq) became a key method of quantitatively measuring the transcriptome. In this study, we compared the detection of genes by each of the transcriptome analysis methods: cDNA array, quantitative RT-PCR, and RNA-seq. As expected, we found differences in the gene expression profiles of the aforementioned techniques. Here, we present selected genes that exemplarily demonstrate the observed differences and calculations to reveal that a strong RNA secondary structure, as well as sample preparation, can affect RNA-seq. In summary, this study addresses an important issue with a strong impact on gene expression analysis in general. Therefore, we suggest that these findings need to be considered when dealing with data from transcriptome analyses.
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Texto completo:
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Bases de dados:
MEDLINE
Assunto principal:
Análise de Sequência de RNA
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Reação em Cadeia da Polimerase Via Transcriptase Reversa
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Análise de Sequência com Séries de Oligonucleotídeos
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Perfilação da Expressão Gênica
Limite:
Humans
Idioma:
En
Revista:
Int J Mol Sci
Ano de publicação:
2021
Tipo de documento:
Article
País de afiliação:
Alemanha