A hybrid RNA FISH immunofluorescence protocol on Drosophila polytene chromosomes.

ABSTRACT:

Objectives

Investigating protein-DNA interactions is imperative to understanding fundamental concepts such as cell growth, differentiation, and cell development in many systems. Sequencing techniques such as ChIP-seq can yield genome-wide DNA binding profiles of transcription factors; however this assay can be expensive, time-consuming, may not be informative for repetitive regions of the genome, and depend heavily upon antibody suitability. Combining DNA fluorescence in situ hybridization (FISH) with immunofluorescence (IF) is a quicker and inexpensive approach which has historically been used to investigate protein-DNA interactions in individual nuclei. However, these assays are sometimes incompatible due to the required denaturation step in DNA FISH that can alter protein epitop

SUBMITTER: Gilbonio HE 

PROVIDER: S-EPMC10486132 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature