癌变·畸变·突变 ›› 1998, Vol. 10 ›› Issue (5): 278-281.doi: 10.3969/j.issn.1004-616x.1998.05.003

• 论著 • 上一篇    下一篇

CHL 细胞DNA 解螺旋荧光分析法实验参数的选择及其应用

薛丽君　金中初   

  1. 浙江医学大学病理生理教研室　杭州　310031
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1998-09-30 发布日期:1998-09-30

THE SELECTION AND THE APPLICATION OF EXPERIMENTAL PARAMETERS ABOUT FLUORMETRY ASSAY OF DNA UNWINDINGON CHL CELLS

Xue Lijun , J in Zhoungchu   

  1. Dept . of Pathophysiology , Zhejiang Medical University , Hangz hou 　310031
  • Received:1900-01-01 Revised:1900-01-01 Online:1998-09-30 Published:1998-09-30

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摘要： 为确定CHL 细胞为测试靶细胞时FADU 的较佳实验参数,采用变化解旋温度及解旋液碱浓度,并根据Birnboim 等的判断标准,结果表明解旋温度先0 ℃30min 后15 ℃60min ;解旋液NaOH 浓度014N 效果较佳。并以此为条件把该法应用于检测过氧化氢(H2O2) 诱导的DNA 过氧化损伤- 适应及抗过氧化作用,结果证明了所选用参数的可靠性。

关键词: CHL 细胞, DNA 解螺旋荧光分析法, 解旋温度, 解旋液碱浓度, 过氧化适应, 抗过氧化

Abstract: To make certaion stable experimental parameters about FADU in detection of CHL DNA breaks , temperature of DNA unwinding and concent ration of unwinding alkaline solution NaOH were modified through FADU on CHL cells. According to Birnboim’s criterion , unwinding temperature first 0 ℃/ 30 min →then 5 ℃/ 60 min and NaOH 0. 4N were selected as better conditions. The reliability of these experimental parameters was established by detection of peroxidative adaptive response and antiperoxidation on CHL DNA breaks induced by H2O2 with the improved FADU.

Key words: CHL cells, FADU, temperature of unwinding, concent ration of unwinding alkaline solution, peroxidative adaptation, antiperoxidation

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