Validation of Endogenous Control Genes for Gene Expression Studies on Human Ocular Surface Epithelium
2011

Identifying Stable Control Genes for Eye Tissue Studies

Sample size: 6 publication Evidence: high

Author Information

Author(s): Bina Kulkarni, Imran Mohammed, Andrew Hopkinson, Harminder Singh Dua

Primary Institution: Queen's Medical Centre, Nottingham, United Kingdom

Hypothesis

To evaluate a panel of ten known endogenous control genes for their stability in human ocular surface epithelial regions.

Conclusion

The study identified stable endogenous control genes for effective normalization in qPCR studies of human ocular surface epithelium.

Supporting Evidence

  • The expression stability of ten known endogenous control genes was evaluated across different ocular surface regions.
  • GeNorm analysis identified PPIA-RPLP0 as the most stable gene pair for normalization.
  • Stable control genes are crucial for accurate gene expression analysis in qPCR studies.

Takeaway

Researchers wanted to find reliable genes to help measure other genes in eye tissue samples, and they found two pairs of genes that work best.

Methodology

The study used laser microdissection to collect RNA from different regions of human cadaver eyes and analyzed gene expression stability using qPCR and geNorm software.

Limitations

The RNA yield from laser microdissected samples was minimal, which may limit the profiling of multiple genes.

Participant Demographics

Cadaver eyes from donors aged 20 to 70 years, of either sex, with undamaged ocular surface epithelium.

Digital Object Identifier (DOI)

10.1371/journal.pone.0022301

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