介绍一种不加盖玻片的核酸原位杂交改良法

A MODIFIED METHOD FOR HYBRIDIZATION IN SITU

核酸原位杂交程序中探针和靶基因的加热变性和杂交为整个程序中最重要的环节,目前国内外最常用的方法是在切片上滴加DNA探针溶液后用盖玻片复盖组织,并加胶泥或其他封固剂封闭玻片四周,变性杂交后需拆除盖玻片,这一操作过程易损坏组织切片或出现干片、脱片。作者针对上述情况,对传统的核酸原位杂交方法中的部分程序加以改良,采用不加盖玻片直接在蒸汽中变性方法,经多次实验证明此方法稳定、可靠、简便,结果满意。

The most critical step in the procedure of hybridization in situ is denaturation by the heating and hybridization aof the probe and the target gene. The conventional method for denaturation includes covering the probe solution with a coverslip, the edge of which is sealed by clay or other adhesive, and then heated. The tissue sections are often damaged, dried or dislo-ged in the process of this step. In this paper a modified method for the step of denaturation of hybridization in situ is introduced. The probe solution is directly heated in a steam bath with no coverslip. The modified method is stable, simple, and reliable, and the effect is satisfactory in our experiences.