Identification of DNA polymorphism induced by X-ray and UV irradiation in plant cells
1999
Kuroda, S. (National Inst. of Agrobiological Resources, Tsukuba, Ibaraki (Japan)) | Yano, H. | Koga Ban, Y. | Tabei, Y. | Takaiwa, F. | Kayano, T. | Tanaka, H.
Angelonia protoplasts were irradiated with X-rays or UV to obtain a half survival rate. To evaluate the DNA damage, total genomic DNA was prepared from protoplasts irradiated with X-rays or UV and nonirradiated control protoplasts, respectively. An aliquot of DNA was subjected to polymerase chain reaction (PCR) with a decamer arbitrary primer in the presence of radiolabeled nucleotides under annealing conditions of low stringency. Randomly amplified PCR products were analyzed using a denaturing polyacrylamide gel. All the DNA bands detected by autoradiography were compared. The number of DNA bands detected in one lane was approximately - 150. The results indicate that the arbitrarily amplified PCR method is very sensitive for monitoring the DNA polymorphism caused by X-ray and UV irradiation. This novel procedure, designated as RI (radioisotope incorporated)-RAPD, could be applied to detect precise DNA polymorphism in plant cells
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