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《Acta Academiae Medicine Militaris Tertiae》 2001-06
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Detection of genomic variation of melanoma cell strain B16 with amplified fragment length polymorphism in C57 mice

LIANG Zhi yong, SHI Jing quan, WEI Hong, WU Feng chun, LIU Li mei, LI Shu rong, HE Guang you (Research Institute of Pathology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China)  
Objective To detect the genetic variant of melanoma cell B16 in C57 mice with amplified fragment length polymorphism (AFLP). Methods Melanoma B16 cells were inoculated into C57 mice after being cultured for 3 to 4 passages. Genomic DNA from melanoma samples and corresponding normal tissue were digested with restriction endonuclease, ligated with specific adapter, and amplified with 20 primers. PCR products were electrophoretically separated in agarose gel and banding profiles were visualized with ethidium bromide staining. Results Clear banding profiles were obtained through the 20 primers and the amplified products showed no difference between those from B16 cell line and from implanted tumor. The amplified products produced by 17 primers were monomorphic, while those by the other 3 primers revealed changes when the products from B16 melanoma AFLP were compared with those from normal tissue DNA. The alterations in tumor DNA included the loss of a normal band and appearance of a new band. Conclusion The occurrence of melanoma might be associated with genetic genomic instability.
【Fund】: 国家自然科学基金资助项目 (3 960 0 0 79;;3 990 0 1 73 ) ;; 全军“九五”医 药卫生科研基金资助项目 (96M0 83 ) ; ; 重庆市攻关项目
【CateGory Index】: R730.23
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