Korean journal of ophthalmology : KJO · 2026
RNA Isolation and qPCR Analysis from Rat Corneal Tissue Following Alkali Burn Injury.
СтатьяНазвание и аннотация: Перевод в очереди
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Yun Da Hyeon, Na Kyung Sun
Аннотация
PURPOSE: To establish a practical protocol for RNA isolation and quantitative real-time polymerase chain reaction (qPCR) analysis using rat corneal tissue following alkali burn injury.
METHODS: Corneal alkali burn injury was induced in rats using a 4-mm filter, paper soaked in 1N NaOH applied to the central cornea for 40 seconds, followed by saline irrigation. Corneal tissues were collected 7 days after injury, and total RNA was isolated using a TRIzol-based protocol optimized for individual corneal samples. RNA concentration and purity were evaluated using spectrophotometry and agarose gel electrophoresis. Complementary DNA was synthesized for qPCR analysis of inflammation- and angiogenesis-related genes.
RESULTS: The optimized protocol enabled successful RNA isolation from individual rat corneal tissues without sample pooling. RNA quality was suitable for downstream qPCR analysis. Expression levels of Vegfa, Mmp9, and Il-6 were successfully detected and showed expected alterations in alkali-burned corneas.
CONCLUSION: This protocol provides a practical and reproducible method for RNA isolation and qPCR analysis in rat corneal alkali burn models and may facilitate molecular investigations of ocular surface inflammation and wound healing.
METHODS: Corneal alkali burn injury was induced in rats using a 4-mm filter, paper soaked in 1N NaOH applied to the central cornea for 40 seconds, followed by saline irrigation. Corneal tissues were collected 7 days after injury, and total RNA was isolated using a TRIzol-based protocol optimized for individual corneal samples. RNA concentration and purity were evaluated using spectrophotometry and agarose gel electrophoresis. Complementary DNA was synthesized for qPCR analysis of inflammation- and angiogenesis-related genes.
RESULTS: The optimized protocol enabled successful RNA isolation from individual rat corneal tissues without sample pooling. RNA quality was suitable for downstream qPCR analysis. Expression levels of Vegfa, Mmp9, and Il-6 were successfully detected and showed expected alterations in alkali-burned corneas.
CONCLUSION: This protocol provides a practical and reproducible method for RNA isolation and qPCR analysis in rat corneal alkali burn models and may facilitate molecular investigations of ocular surface inflammation and wound healing.