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Selection of Suitable Reference Genes for Normalization of Quantitative Real-Time Polymerase Chain Reaction in Human Cartilage Endplate of the Lumbar Spine

The quantitative real-time polymerase chain reaction (qRT-PCR) is one of the most widely used methods to study gene expression profiles, and it requires appropriate normalization for accurate and reliable results. Although several genes are commonly used as reference genes (such as GAPDH, ACTB, and... Full description

Journal Title: PLoS One Feb 2014, Vol.9(2), p.e88892
Main Author: Zhou, Zhi-Jie
Other Authors: Jian-Feng, Zhang , Xia, Ping , Ji-Ying, Wang , Chen, Shuai , Xiang-Qian, Fang , Shun-Wu, Fan
Format: Electronic Article Electronic Article
Language: English
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ID: E-ISSN: 19326203 ; DOI: 10.1371/journal.pone.0088892
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title: Selection of Suitable Reference Genes for Normalization of Quantitative Real-Time Polymerase Chain Reaction in Human Cartilage Endplate of the Lumbar Spine
format: Article
creator:
  • Zhou, Zhi-Jie
  • Jian-Feng, Zhang
  • Xia, Ping
  • Ji-Ying, Wang
  • Chen, Shuai
  • Xiang-Qian, Fang
  • Shun-Wu, Fan
subjects:
  • China
  • Cartilage
  • Rrna 18s
  • Clinical Medicine
  • Genes
  • Nuclear Magnetic Resonance–NMR
  • Spine (Lumbar)
  • Cartilage
  • Hospitals
  • Polymerase Chain Reaction
  • Genomes
  • Spine
  • Polymerase Chain Reaction
  • Computer Simulation
  • Gene Expression
  • Medicine
  • Arthritis
  • Polymerase Chain Reaction
  • Surgery
ispartof: PLoS One, Feb 2014, Vol.9(2), p.e88892
description: The quantitative real-time polymerase chain reaction (qRT-PCR) is one of the most widely used methods to study gene expression profiles, and it requires appropriate normalization for accurate and reliable results. Although several genes are commonly used as reference genes (such as GAPDH, ACTB, and 18S rRNA), they are also regulated and can be expressed at varying levels. In this study, we evaluated twelve well-known reference genes to identify the most suitable housekeeping gene for normalization of qRT-PCR in human lumbar vertebral endplate with Modic changes, by using the geNorm, NormFinder, and BestKeeper algorithms. Our results showed that the rarely-used SDHA was the most stable single reference gene, and a combination of three, SDHA, B2M, and LDHA, was the most suitable gene set for normalization in all samples. In addition, the commonly-used genes, GAPDH, ACTB and 18S rRNA, were all inappropriate as internal standards. The rankings of reference genes for the three types of Modic...
language: eng
source:
identifier: E-ISSN: 19326203 ; DOI: 10.1371/journal.pone.0088892
fulltext: fulltext_linktorsrc
issn:
  • 19326203
  • 1932-6203
url: Link


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titleSelection of Suitable Reference Genes for Normalization of Quantitative Real-Time Polymerase Chain Reaction in Human Cartilage Endplate of the Lumbar Spine
creatorZhou, Zhi-Jie ; Jian-Feng, Zhang ; Xia, Ping ; Ji-Ying, Wang ; Chen, Shuai ; Xiang-Qian, Fang ; Shun-Wu, Fan
ispartofPLoS One, Feb 2014, Vol.9(2), p.e88892
identifierE-ISSN: 19326203 ; DOI: 10.1371/journal.pone.0088892
subjectChina ; Cartilage ; Rrna 18s ; Clinical Medicine ; Genes ; Nuclear Magnetic Resonance–NMR ; Spine (Lumbar) ; Cartilage ; Hospitals ; Polymerase Chain Reaction ; Genomes ; Spine ; Polymerase Chain Reaction ; Computer Simulation ; Gene Expression ; Medicine ; Arthritis ; Polymerase Chain Reaction ; Surgery
descriptionThe quantitative real-time polymerase chain reaction (qRT-PCR) is one of the most widely used methods to study gene expression profiles, and it requires appropriate normalization for accurate and reliable results. Although several genes are commonly used as reference genes (such as GAPDH, ACTB, and 18S rRNA), they are also regulated and can be expressed at varying levels. In this study, we evaluated twelve well-known reference genes to identify the most suitable housekeeping gene for normalization of qRT-PCR in human lumbar vertebral endplate with Modic changes, by using the geNorm, NormFinder, and BestKeeper algorithms. Our results showed that the rarely-used SDHA was the most stable single reference gene, and a combination of three, SDHA, B2M, and LDHA, was the most suitable gene set for normalization in all samples. In addition, the commonly-used genes, GAPDH, ACTB and 18S rRNA, were all inappropriate as internal standards. The rankings of reference genes for the three types of Modic...
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titleSelection of Suitable Reference Genes for Normalization of Quantitative Real-Time Polymerase Chain Reaction in Human Cartilage Endplate of the Lumbar Spine
descriptionThe quantitative real-time polymerase chain reaction (qRT-PCR) is one of the most widely used methods to study gene expression profiles, and it requires appropriate normalization for accurate and reliable results. Although several genes are commonly used as reference genes (such as GAPDH, ACTB, and 18S rRNA), they are also regulated and can be expressed at varying levels. In this study, we evaluated twelve well-known reference genes to identify the most suitable housekeeping gene for normalization of qRT-PCR in human lumbar vertebral endplate with Modic changes, by using the geNorm, NormFinder, and BestKeeper algorithms. Our results showed that the rarely-used SDHA was the most stable single reference gene, and a combination of three, SDHA, B2M, and LDHA, was the most suitable gene set for normalization in all samples. In addition, the commonly-used genes, GAPDH, ACTB and 18S rRNA, were all inappropriate as internal standards. The rankings of reference genes for the three types of Modic...
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titleSelection of Suitable Reference Genes for Normalization of Quantitative Real-Time Polymerase Chain Reaction in Human Cartilage Endplate of the Lumbar Spine
authorZhou, Zhi-Jie ; Jian-Feng, Zhang ; Xia, Ping ; Ji-Ying, Wang ; Chen, Shuai ; Xiang-Qian, Fang ; Shun-Wu, Fan
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abstractThe quantitative real-time polymerase chain reaction (qRT-PCR) is one of the most widely used methods to study gene expression profiles, and it requires appropriate normalization for accurate and reliable results. Although several genes are commonly used as reference genes (such as GAPDH, ACTB, and 18S rRNA), they are also regulated and can be expressed at varying levels. In this study, we evaluated twelve well-known reference genes to identify the most suitable housekeeping gene for normalization of qRT-PCR in human lumbar vertebral endplate with Modic changes, by using the geNorm, NormFinder, and BestKeeper algorithms. Our results showed that the rarely-used SDHA was the most stable single reference gene, and a combination of three, SDHA, B2M, and LDHA, was the most suitable gene set for normalization in all samples. In addition, the commonly-used genes, GAPDH, ACTB and 18S rRNA, were all inappropriate as internal standards. The rankings of reference genes for the three types of Modic...
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