Watters Amanda D, Stacey Michael W, Bartlett John
University Dept of Surgery, Glasgow Royal Infirmary, Scotland, UK.
Mol Biotechnol. 2002 Mar;20(3):257-60. doi: 10.1385/mb:20:3:257.
Nick translation is used to label DNA and RNA to produce probes for in situ hybridization and Northern and Southern blotting. Fluorescence in situ hybridization (FISH) is a widely applied technique used to determine chromosomal and genetic anomalies in many biological samples. Initially the technique was applied to metaphase preparations, but the usefulness of detecting genetic anomalies in solid tumors in situ has resulted in the development of modified protocols. Formalin fixed paraffin processed tissue sections present novel challenges when applying FISH; the probes must be small (between 200 and 600 base pairs) and pretreatment is necessary before the probes can be applied to tissue sections, to promote probe access to target DNA. Here we report on a modification of a nick translation method to produce a probe that can reliably be used with FISH in paraffin processed tissue sections.
缺口平移法用于标记DNA和RNA，以制备用于原位杂交以及Northern和Southern印迹的探针。荧光原位杂交（FISH）是一种广泛应用的技术，用于确定许多生物样品中的染色体和基因异常。最初，该技术应用于中期染色体标本制备，但在实体瘤原位检测基因异常的实用性促使了改良方案的发展。当应用FISH时，经福尔马林固定、石蜡包埋处理的组织切片带来了新的挑战；探针必须小（200至600个碱基对之间），并且在将探针应用于组织切片之前需要进行预处理，以促进探针与靶DNA的结合。在此，我们报告了一种缺口平移法的改良方法，以产生一种可可靠地用于经石蜡处理的组织切片的FISH探针。
