Total RNA extraction and single-strand cDNA synthesis

DK Dong Ju Kim
MJ Mi-Young Jung
HP Ha-Jin Pak
JP Joo-Hee Park
MK Martha Kim
RC Roy S. Chuck
CP Choul Yong Park
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After exposure to the HAM, primary corneal epithelial cells were harvested, ground up with a grinder in 5 volumes of TRIZOL reagent (Molecular Research Center, Inc.), and stored at 80 °C until use. Total RNA was isolated from harvested primary corneal epithelial cells according to the manufacturer’s instructions. Total RNA quality and quantity was confirmed using 1% agarose gels with RedSafe (Intron Biotechnology) staining, a UV transilluminator (Vilber Lourmat, TFX-20), and a Nano drop spectrophotometer (ACTGene Inc., ASP-2680). Single-strand cDNA was synthesized from 1 μg of total RNA using random primers for reverse transcription (Toyobo, ReverTra Ace qPCR RT Master Mix, Japan).

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