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固定剂对染色质结构和DNA检测的影响。

Effect of fixative on chromatin structure and DNA detection.

作者信息

Falconi Mirella, Teti Gabriella, Zago Michela, Pelotti Susi, Gobbi Pietro, Breschi Lorenzo, Mazzotti Giovanni

机构信息

Department of Anatomical Sciences, University of Bologna, Italy.

出版信息

Microsc Res Tech. 2007 Jul;70(7):599-606. doi: 10.1002/jemt.20440.

Abstract

In this study, we analyzed the chromatin ultrastructure in interphase cells after different chemical fixations. In light of the fact that there is little information regarding the fixation of biological samples in combination with molecular biology methods (such as DNA extraction and in situ hybridization methods) we analyzed the ultrastructure of chromatin in interphase cells fixed with different fixatives and tested under the same conditions for both DNA extraction and in situ hybridization. The results showed that, among the different combinations and concentrations we analyzed, the solution of 4% paraformaldehyde/0.1% glutaraldehyde was the best compromise in order to achieve a well-preserved morphology, successful DNA extraction, and specific signaling of in situ hybridization, suggesting a low interference of this fixative with the chromatin organization.

摘要

在本研究中,我们分析了不同化学固定后间期细胞中的染色质超微结构。鉴于结合分子生物学方法(如DNA提取和原位杂交方法)对生物样品进行固定的信息较少,我们分析了用不同固定剂固定的间期细胞中染色质的超微结构,并在相同条件下对DNA提取和原位杂交进行了测试。结果表明,在我们分析的不同组合和浓度中,4%多聚甲醛/0.1%戊二醛溶液是实现形态良好保存、DNA成功提取和原位杂交特异性信号的最佳折衷方案,表明该固定剂对染色质组织的干扰较小。

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