多层电泳槽在彗星试验中的应用

Application of multilayer electrophoresis tank in comet assay

目的本研究拟通过彗星试验来验证多层电泳槽的实用性,从而推进彗星试验的规范化。方法从10只SD大鼠中分别提取淋巴细胞,各制作6张彗星试验载玻片,其中每只动物的一半载玻片用30%H2O2处理作为阳性组,另一半未处理的作为阴性组;分别用单层电泳和多层电泳开展彗星试验,并分析彗星试验结果。结果 30%H2 O2导致淋巴细胞DNA损伤;单层电泳和多层电泳的阳性组相比差异有显著性;多层电泳的不同层之间差异无显著性。结论彗星试验同时进行电泳比分次电泳更有可比性;多层电泳槽能提高彗星试验单次电泳的样品数量,有利于彗星试验的规范化。

Objective In order to standardize the comet assay,a multilayer electrophoresis tank was invented and validated.Methods Lymphocytes were extracted from ten SD rats.Ten slides were prepared for the lymphocytes of each rat for comet assay.One half slides were treated with 30% H2 O2,and the other untreated slides were used as negative control.Comet assays on single layer-tank or multi-layer tank were used for comparing the assay results.Results The DNA of lymphocytes was damaged significantly by 30% H2 O2.There were a significant differences of results between single-layer tank and multilayer tank,but no significant differences of results among different layers of multilayer tank.Conclusion Comparing with the single-layer tank,more samples could be analyzed in multilayer tank,and the comparability of results for comet assay were improved.Multilayer tank would be helpful to standardize comet assays.