How do I calculate the hybridization temperature for my DNA-FISH probe?

31 March 2021 1 4K Report

Hi everyone,

I have a DNA-FISH probe of 3 kb in length. I'm used to use very long probes (>100 kb) and hybridize them overnight at 37 C. Now I have to perform a FISH protocol using this 3 kb probe and I don't know which temperature to use and for how long to do the hybridization step. Can somebody help me with that?

José Francisco Carrasco

I can't help you directly but maybe the following paper will be useful:

Sílvia Fontenete,Nuno Guimarães,Jesper Wengel & Nuno Filipe Azevedo (2014). Prediction of melting temperatures in fluorescence in situ hybridization (FISH) procedures using thermodynamic models, Critical Reviews in Biotechnology, 36(3): 566-577.

https://doi.org/10.3109/07388551.2014.993589

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