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Sensitive and specific method for detecting G protein–coupled receptor mRNAs

Nature Methods volume 4, pages 35–37 (2007)Cite this article

Abstract

G protein–coupled receptors (GPCRs) mediate effects of extracellular signaling molecules in all the body's cells. These receptors are encoded by scarce mRNAs; therefore, detecting their transcripts with conventional microarrays is difficult. We present a method based on multiplex PCR and array detection of amplicons to assay GPCR gene expression with as little as 1 μg of total RNA, and using it, we profiled three human bone marrow stromal cell (BMSC) lines.

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Figure 1: GPCR transcript detection.
Figure 2: Venn diagrams illustrating GPCR expression profiles of BMSCs.

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Acknowledgements

This project was funded in part with Federal funds from the US National Cancer Institute, under Contract number N01-C0-12400, by the Intramural Research Programs of the National Institute of Mental Health, the National Institute of Neurological Disorder and Stroke, and the National Institute of Dental and Craniofacial Research, US National Institutes of Health, Deutsche Forschungsgemeinschaft Ha3423/2-1 and the J. Craig Venter Foundation.

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Authors and Affiliations

  1. Department of Health and Human Services, Craniofacial and Skeletal Diseases Branch, National Institute of Dental and Craniofacial Research, Bethesda, 20892, Maryland, USA

    Arne Hansen, Natasha Cherman, Sergei A Kuznetsov, Pamela G Robey & Eva Mezey

  2. Department of Health and Human Services, Cancer Genetics Branch, National Human Genome Research Institute, Bethesda, 20892, Maryland, USA

    Yidong Chen

  3. Department of Health and Human Services, Laboratory of Neurochemistry, National Institute of Neurological Disorder and Stroke, Bethesda, 20892, Maryland, USA

    Arne Hansen & Eva Mezey

  4. Department of Health and Human Services, National Institute of Mental Health, National Institutes of Health, Bethesda, 20892, Maryland, USA

    Quang N Phan, Zhi-Qing Qi, Charlie C Xiang, Miklos Palkovits & Michael J Brownstein

  5. The Institute for Genomic Research, Rockville, 20850, Maryland, USA

    Jason M Inman

  6. J. Craig Venter Institute, Rockville, 20850, Maryland, USA

    Quang N Phan, Charlie C Xiang & Michael J Brownstein

Authors
  1. Arne Hansen
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  2. Yidong Chen
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  8. Natasha Cherman
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  9. Sergei A Kuznetsov
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  10. Pamela G Robey
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  11. Eva Mezey
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  12. Michael J Brownstein
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Corresponding authors

Correspondence to Arne Hansen or Michael J Brownstein.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Table 1

GPCRs expressed in human BMSCs.

Supplementary Table 2

Quantitative PCR analysis of receptors that are inconsistently expressed in BMSC2.

Supplementary Table 3

GPCRs expressed in human liver, kidney, ovary, hippocampus and cerebral (temporal) cortex.

Supplementary Table 4

List of GPCR oligonucleotide probes.

Supplementary Table 5

List of GPCR PCR primers.

Supplementary Note 1

Determination of sensitivity and specificity.

Supplementary Note 2

Evaluation by quantitative PCR.

Supplementary Note 3

Data analysis.

Supplementary Methods

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Hansen, A., Chen, Y., Inman, J. et al. Sensitive and specific method for detecting G protein–coupled receptor mRNAs. Nat Methods 4, 35–37 (2007). https://doi.org/10.1038/nmeth977

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