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  • High-throughput analysis of gene expression on tissue sections by in situ hybridization.

High-throughput analysis of gene expression on tissue sections by in situ hybridization.

Methods (San Diego, Calif.) (2010-12-28)
Gregor Eichele, Graciana Diez-Roux
ABSTRACT

Genome-scale sequencing projects, high-throughput RNAi screens, systematic gene targeting, and system-biology-based network predictions all depend on a validation of biological significance in order to understand the relevance of a particular finding. Such validation, for the most part, rests on low-throughput technologies. This article provides protocols that, in combination with suitable instrumentation, make possible a semi-automated analysis of gene expression on tissue sections by means of in situ hybridization. Knowledge of gene expression localization has the potential to aid, and thereby accelerate, the validation of gene functions.

MATERIALS
Product Number
Brand
Product Description

Roche
BCIP, solution, >95% (HPLC), pkg of 3 mL (150 mg)
Sigma-Aldrich
Sheep Serum, USA origin, sterile-filtered, suitable for cell culture
Sigma-Aldrich
(−)-Tetramisole hydrochloride, ≥99% (GC)
Roche
Anti-Fluorescein-POD, Fab fragments, from sheep
Roche
Anti-Digoxigenin-POD, Fab fragments, from sheep
Roche
Blocking Reagent, For nucleic acid hybridization and detection
Roche
NBT, 4-Nitro blue tetrazolium chloride, solution