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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
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A PCR-based Genotyping Method to Distinguish Between Wild-type and Ornamental Varieties of Imperata cylindrica
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[Reference gene screening for Real-time quantitative PCR in Polygonum multiflorum].

Hao Wang1, Qi-Zhong Cai1, Lu Liu1

  • 1Guangdong Engineering Research Center of Good Agricultural Practice, Comprehensive Development for Cantonese Medicinal Materials,Guangzhou Comprehensive Experimental Station of National Industrial Technology System for Chinese Materia Medica,Key Laboratory of State Administration of Traditional Chinese Medicine for Production, Development of Cantonese Medicinal Materials, School of Traditional Chinese Medicine, Guangdong Pharmaceutical University Guangzhou 510006, China.

Zhongguo Zhong Yao Za Zhi = Zhongguo Zhongyao Zazhi = China Journal of Chinese Materia Medica
|March 1, 2021
PubMed
Summary

The PP2A gene is the most stable reference gene for analyzing gene expression in Polygonum multiflorum tissues. This finding ensures reliable gene expression analysis in P. multiflorum research.

Keywords:
Polygonum multiflorumReal-time quantitative PCRreference gene

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Area of Science:

  • Molecular Biology
  • Plant Science
  • Genetics

Background:

  • Accurate gene expression analysis in Polygonum multiflorum requires validated reference genes.
  • Transcriptome data identified five candidate reference genes: Actin, GAPDH, SAND, PP2A, and TIP41.

Purpose of the Study:

  • To evaluate the expression stability of five candidate reference genes in different tissues of P. multiflorum.
  • To identify the most suitable reference gene for quantitative gene expression studies in P. multiflorum.

Main Methods:

  • Candidate reference gene primers were designed.
  • Real-time quantitative PCR (RT-qPCR) was used to analyze gene expression.
  • Expression stability was assessed using geNorm, NormFinder, BestKeeper, Delta CT, and RefFinder algorithms.

Main Results:

  • Significant variations in expression levels and stability were observed among candidate genes across different tissues.
  • Actin and GAPDH showed high expression levels; SAND, PP2A, and TIP41 showed lower levels.
  • PP2A and GAPDH were most stable according to geNorm; PP2A was most stable by NormFinder and Delta CT; Actin was most stable by BestKeeper.
  • RefFinder analysis indicated PP2A as the most stable, followed by GAPDH, Actin, TIP41, and SAND.

Conclusions:

  • The PP2A gene is identified as the most stable and ideal reference gene for P. multiflorum gene expression analysis.
  • Validated reference genes are crucial for reliable gene expression profiling in plant research.