A Widely Applicable Urea-based Fluorescent/Colorimetric mRNA in situ Hybridization Protocol.

ABSTRACT: In situ hybridization methods are routinely employed to detect nucleic acid sequences, allowing to localize gene expression or to study chromosomal organization in their native context. These methods rely on the pairwise binding of a labeled probe to the target endogenous nucleic acid sequence-the hybridization step, followed by detection of annealed sequences by means of fluorescent or colorimetric reactions. Successful hybridization requires permeabilization of tissues, followed by denaturation of nucleic acids strands, which is usually carried out in a formamide-based buffer and at high temperatures. Such reaction conditions, besides posing a health hazard (both concerning manipulation and waste disposal), can be excessively harsh for the delicate tissues of some species or devel

SUBMITTER: Sinigaglia C 

PROVIDER: S-EPMC7854207 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature