Methods

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    Kjetil Hodne

We use a combination of molecular biology, cellular physiology and genomics tools.

Methods

Molecular biology

A fundamental part of our research is gene expression profiling. In this work we use standard quantitative PCR. In addition, we have specialized our research in measuring low amounts of RNA down to single cells using digital droplet PCR. For detection and quantification of proteins and hormones we use western blot, ELISA and RIA techniques. Finally, for in vivo labeling of cell types we have developed several transgenic animals, and more recently we are now using CRISPR/Cas9 technology for gene specific knockout.

 

Cellular physiology

Following gene expression profiling we combine a variety of techniques to elucidate fundamental question regarding cellular physiology. Multicolor in situ hybridization and immunofluorescence techniques in combination with confocal imaging are the main tools for characterizing the different cells. Normal histology is also used for cell labeling and investigating structure and organization.  Functional studies are performed using electrophysiology and calcium imaging techniques. The electrophysiological experiments are performed using patch-clamping. The patch-clamp setup also includes equipment for uncaging experiments. Uncaging is a powerful method that allows us to activate different molecules like Ca2+, ATP, glutamate and IP3 using a UV laser. Uncaging allow us to manipulate cells and at the same time record the response.

 

Genomics

We use Illumina RNA-seq for gene expression profiling, as well as single-cell RNA-seq (based on the 10x Genomics platform) for cell type identification. In addition, we use Oxford Nanopore technology for sequencing fish genomes, which we analyze using a variety of comparative genomics approaches.

Published 18. November 2018 - 11:24 - Updated 22. November 2018 - 15:15