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Open Access Highly Accessed Research article

Evaluation of putative reference genes for gene expression normalization in soybean by quantitative real-time RT-PCR

Ruibo Hu, Chengming Fan, Hongyu Li, Qingzhu Zhang and Yong-Fu Fu*

Author Affiliations

Institute of Crop Science, National Key Facility of Crop Gene Resource and Genetic Improvement, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China

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BMC Molecular Biology 2009, 10:93  doi:10.1186/1471-2199-10-93

Published: 28 September 2009

Additional files

Additional file 1:

List of reference genes used for gene expression studies in soybean. The list comprises 54 hits from a search (January 2001 to March 2009) of PubMed, using "soybean" and "gene expression" as keywords.

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Additional file 2:

The transcription profiles of individual reference genes given as absolute Cq values across all samples. The scatter plots show the expression levels of the various reference genes. Values are given in the form of quantification cycle numbers (Cq values).

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Open Data

Additional file 3:

Representative amplification plots and melting curves obtained in the RT-qPCR efficiency test. Four to five ten-fold serial dilutions were plotted against the logarithm of cDNA template concentration. Amplification plots and melting curve images were collected using StepOne software v2.0 (Applied Biosystems).

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Additional file 4:

RT-qPCR primer efficiency plots. Mean quantification cycle (Cq) values of each set of ten-fold serial dilution plotted against the logarithm of cDNA template concentration. The reaction efficiency (E) is given by [10(1/-S)-1] × 100%, where S represents the slope of the linear regression line.

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Additional file 5:

Tissue/organ sample sets used for the analysis of gene expression. See Methods section for details.

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Additional file 6:

Expression profiling of six conventional housekeeping genes, using microarray data derived from Genevestigator. The Meta-Profile Analysis tool was used to produce expression profiling from representative UniGene IDs. No probes available for TUB4.

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Additional file 7:

Expression profiling of seven new reference genes tested from Genevestigator microarray data. The Meta-Profile Analysis tool was used to produce expression profiling from representative UniGene IDs.

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Open Data