Abstract
In situ hybridization in zebrafish embryos frequently suffers from high background signal. Torular yeast RNA is often added to reduce the non-specific binding that leads to the high background signal. Without appropriate preparation, torular yeast RNA can also add background to the staining. This method is an easy and quick way to clean torular RNA for Hybe solution used in in situ hybridization.
Materials and Reagents
Equipment
Procedure
Acknowledgments
This protocol was modified from the original protocol by Caroline Burns at the Len Zon lab, Boston Children’s Hospital, Boston, USA and supported by NIH grant R01 HL04880-21.
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Regular phenol works, like the one from invitrogen, cat #: 15513-039.
Yes, ethanol (ethyl alcohol, 200 proof from Pharmaco-AAPER) is used to precipitate the yeast RNA at steps 8 and 11.