Abstract
In situ hybridization is an effective method to examine the expression level and location of a gene of interest in tissues or cells. To do this, RNA can be labeled with digoxigenin-UTP (DIG) by in vitro transcription with SP6 and T7 RNA polymerase. The method provided in this protocol is a detailed description of synthesizing an antisense DIG-labeled RNA probe for in situ hybridization using the DIG RNA labeling kit from Roche.
Materials and Reagents
Equipment
Procedure
Notes
I usually synthesize 3-4 separate dig probe reactions of the same probe and add all of them to the same G50 column. This way you have made quite a bit of probe using only 1 column.
Acknowledgments
This protocol was developed in the Michael Granato Lab at University of Pennsylvania, Philadelphia, USA, and this work was supported by NIH grant R01HD037975.
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