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Published on: June 30, 2023
RNA Isolation and qPCR Analysis from Rat Corneal Tissue Following Alkali Burn Injury.
1Department of Ophthalmology and Visual Science, Yeouido St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
Korean Journal of Ophthalmology : KJO
|July 8, 2026
Summary
This study presents a new method for isolating RNA from rat corneas after alkali burns, enabling gene expression analysis via quantitative real-time polymerase chain reaction (qPCR). The protocol successfully detects key inflammation and angiogenesis genes, aiding ocular surface research.
Area of Science:
- Ophthalmology
- Molecular Biology
- Biochemistry
Background:
- Corneal alkali burns are severe injuries requiring effective molecular analysis for treatment development.
- Accurate RNA isolation is crucial for studying gene expression changes in ocular surface diseases.
Purpose of the Study:
- To develop and validate a practical protocol for RNA isolation from individual rat corneas post-alkali burn.
- To enable quantitative real-time polymerase chain reaction (qPCR) analysis of gene expression in corneal tissues.
Main Methods:
- Rat corneal alkali burn model induced with sodium hydroxide (NaOH).
- TRIzol-based RNA isolation optimized for single corneal samples.
- RNA quality assessed by spectrophotometry and gel electrophoresis.
- Complementary DNA synthesis for qPCR analysis of inflammation and angiogenesis genes.
Main Results:
- Successful RNA isolation from individual rat corneas without pooling.
- High-quality RNA suitable for downstream qPCR.
- Detection of altered expression for key genes (Vegfa, Mmp9, Il-6) in burned corneas.
Conclusions:
- The developed protocol is practical and reproducible for RNA isolation and qPCR in rat corneal alkali burn models.
- Facilitates molecular investigation of ocular surface inflammation and wound healing processes.
- Supports research into novel therapeutic strategies for corneal injuries.
