FISH with and Without COT1 DNA

Complex FISH probes comprising large spans of genomic DNA always contain a high amount of dispersed repetitive sequences hampering the visualization of specific signals. To overcome this problem, different approaches have been elaborated that depend on experiment type and probe quality. A classical way to suppress repetitive sequences is to use unlabelled competitor DNA (sheared total genomic DNA or repeated sequences enriched DNA fractions). Here we present two protocols—the first one describes a rapid COT DNA isolation and peculiarities of its use in different FISH experiments, and the second is elaborated for COT-free FISH with complex probes and is based on a special software tool for image enhancement.

Source: Springer protocols feed by Cell Biology - October 20, 2016 Category: Cytology Source Type: news