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Real Time RT-PCR

Real Time RT-PCR

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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Reference gene selection for gene expression studies in lily using quantitative real-time PCR.

M F Zhang¹, Q Liu¹, G X Jia¹

¹Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding, National Engineering Research Center for Floriculture and College of Landscape Architecture, Beijing Forestry University, Beijing, China.

Genetics and Molecular Research : GMR

|May 14, 2016

Summary

This study identifies the most stable reference genes for gene expression analysis in lilies using quantitative real-time polymerase chain reaction (qRT-PCR). TIP, EF, Clathrin, and BHLH are recommended for accurate gene expression studies in lily cultivars.

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Area of Science:

Plant molecular biology
Gene expression analysis
Genomics

Background:

Quantitative real-time polymerase chain reaction (qRT-PCR) is crucial for gene expression analysis.
Selecting stable reference genes is essential for accurate qRT-PCR results.
Reference genes must exhibit consistent expression across different tissues and developmental stages.

Purpose of the Study:

To identify and validate the most stable reference genes for gene expression studies in lily (Lilium x formolongi "Raizan 3").
To provide reliable reference genes for analyzing flowering time and floral development genes in lilies.
To evaluate the expression stability of nine candidate reference genes.

Main Methods:

Digital gene expression technology was used to select nine candidate reference genes.
Gene expression stability was analyzed using three methods: GeNorm, NormFinder, and BestKeeper.
A total of 144 lily samples from various tissues, developmental stages, and hormone treatments were analyzed.

Main Results:

The expression stability of the nine candidate reference genes varied significantly across different sample types.
TIP, EF, Clathrin, and BHLH were identified as the most stable reference genes overall.
The expression of LfFT was analyzed in different lily tissues to validate the selected reference genes.

Conclusions:

TIP, EF, Clathrin, and BHLH are recommended as reliable reference genes for qRT-PCR in lily.
This study provides a valuable resource for researchers studying gene expression in lily cultivars.
Accurate reference gene selection is critical for understanding flowering time and floral development in lilies.