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Report No.

DNA damage evaluation system of the high-LET ion beam using the polymerase chain reaction

Matsuo, Yoichiro*; Izumi, Yoshinobu*; Hase, Yoshihiro; Sakamoto, Ayako; Nozawa, Shigeki; Narumi, Issei; Shimizu, Kikuo*

We have been studying ion beam-induced mutations in budding yeast S288c (${it RAD}$ $$^{+}$$) as a model of eukaryote cell. We report a new method to evaluate DNA lesions caused by high-LET radiation using the polymerase chain reaction (PCR). PCR is one of the most reliable methods for detecting DNA damage as the amplification stops at the site of the damage. In this study, the 804-bp region of ${it URA3}$ gene was amplified by PCR reaction using a specific oligonucleotide primer set. The PCR device adopted was an Eco Real-Time PCR System (Illumina). The percentage of undamaged template DNA was tended to decrease with an increase in absorbed dose of radiation. The higher LET radiations resulted in the higher rate of decrease in undamaged template DNA. This result suggests that different types of lesions are produced on DNA depending on the LET value of radiations.



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