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首页> 外文期刊>Canadian Journal of Pure and Applied Sciences >INCREASED SENSITIVITY OF APOPTOSIS DETECTION USING DIRECT DNA STAINING METHOD AND INTEGRATION OF ACRIDINE ORANGE AS AN ALTERNATIVE SAFER FLUORESCENT DYE IN AGAROSE GEL ELECTROPHORESIS AND MICRONUCLEUS TEST
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INCREASED SENSITIVITY OF APOPTOSIS DETECTION USING DIRECT DNA STAINING METHOD AND INTEGRATION OF ACRIDINE ORANGE AS AN ALTERNATIVE SAFER FLUORESCENT DYE IN AGAROSE GEL ELECTROPHORESIS AND MICRONUCLEUS TEST

机译：直接DNA染色法和AND啶橙的整合作为替代SAFE荧光染料在琼脂糖凝胶电泳和微核试验中凋亡检测的敏感性增加

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Electrophoresis is a common technique used in identification, quantification and purification of nucleic acid fragments. Some troubleshooting faced us in detection of apoptosis through DNA fragmentation assay. In some samples; electrophoretic pattern of extracted DNA has a flam shape nearly like sever necrotic fragmentation. The new method was evaluated in this study to standardize the apoptotic fragmentation detection. The direct incubation of extracted DNA samples which have a flam shape with ethidium bromide (EtBr) and acridine orange (AO) gave better results than the ordinary method of gel staining in apoptotic bands detection which appeared at 180 bp and its multiples on 1.8% agarose gel. Moreover, due to the hazardous effect of EtBr handling, acridine orange was tested in parallel on the same nucleic acid samples as an alternative fluorescent staining dye. Additionally, a novel micronucleus test can be accomplished in parallel with viability investigation by fluorescent dual AO/EtBr method. The obtained results showed obvious micronucleated and other nuclear abnormalities observations. In conclusion, although acridine orange is less sensitive than ethidium bromide staining in case of agarose electrophoresis, its safety, low cost and metachromatic properties should be considered and advisably, acridine orange can be used as an alternative nucleic acids fluorescent dye with increased sensitivity for apoptosis detection using direct incubation method. Acridine orange can also be integrated for micronucleus technique.
机译：电泳是用于鉴定，定量和纯化核酸片段的常用技术。通过DNA片段分析检测细胞凋亡时，我们面临一些疑难解答。在一些样品中;提取的DNA的电泳图样呈火焰状，几乎像严重的坏死碎片一样。在这项研究中评估了新方法，以标准化凋亡片段检测。将具有火焰形状的提取的DNA样品与溴化乙锭（EtBr）和a啶橙（AO）直接温育，在凋亡条带检测中的常规条带染色方法比在180 bp处出现，并且在1.8％琼脂糖上具有倍数时，能获得更好的结果。凝胶。此外，由于处理EtBr的有害影响，在相同的核酸样品上并行测试了cr啶橙，作为荧光染料的替代染料。此外，可以通过荧光双AO / EtBr方法与可行性研究同时完成新的微核试验。获得的结果显示出明显的微核和其他核异常观察。总之，尽管在琼脂糖电泳的情况下a啶橙不如溴化乙锭染色敏感，但应考虑其安全性，低成本和变色特性，建议将a啶橙用作替代的核酸荧光染料，增加对细胞凋亡的敏感性使用直接孵育方法进行检测。 cr啶橙也可以整合到微核技术中。

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