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RISS 인기검색어
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- 주제어 : Neuroblastoma cell line SK-N-BE (검색결과 1 건)
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자기방사능법에 의한 DNA in situ Hybridization방법
장성익,양창헌,김대광,정용욱,최인장,이인환 계명대학교 의과대학 1991 계명의대학술지 Vol.10 No.2
To find out the optimal condition for DNA in situ hybridzation using isotopically labeled DNA probe with the general goal of understanding how localization and organization of N-myc oncogene sequences within the SK-N-BE cell line, present investigation was carried out by ³H-labeled pMF3-SEBI-Nmyc probe in situ hybridization. The results were as follows: Specific activities of 2~5×10??? dpm/㎍ of DNA were gained after separation of the ³H labeled DNA by spindialysis using sepharose CL-6B. The chromosomes and interphase nuclei were hybridization with the labeled probe for 14hr at 41℃ in a hybridization mixture solution: 50% formamide, 2×SSCP(2×SSC, 0.04M NaPO4, pH7.0), 10% dextran sulfate, 100㎍/ml of sonicated salmon sperm DNA and 0.2mg/ml each of bovine serum albumin, Ficoll, and polyvinylphyrrolidone. Autoradiography was performed by using half-strength Sakura NR-M2 emulsion for 7-14 days at 4℃. Silver grains were simultaneously detected on Q-banded chromosomes in the same metaphase. Amplifications of N-myc oncogene were conformed on homogeneous staining regions of chromosome 11 and marker chromosome by statistical analysis. It was impossible with this technique to localize N-myc oncogene within the interphase nucleus of SK-N-BE cell line.
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