Hybridization chain reaction (HCR) protocol for Gastruloids (ESC aggregates) v1

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Abstract

This protocol is for the in situ hybridization chain reaction (HCR) of Gastruloids (3D aggregates of mouse embryonic stem cells). This is an adaptation of the same protocol as applied to mouse tails (Sanchez et al; dx.doi.org/10.17504/protocols.io.7pyhmpw) in turn adapted from the whole-mount mouse embryo protocol available on the website of Molecular Instruments (for details, please follow this link). References Choi, H., Schwarzkopf, M., Fornace, M., Acharya, A., Artavanis, G., & Stegmaier, J. et al. (2018). Third-generationin situhybridization chain reaction: multiplexed, quantitative, sensitive, versatile, robust.Development,145 (12), dev165753. doi: 10.1242/dev.165753 Choi, H., Calvert, C., Husain, N., Huss, D., Barsi, J., & Deverman, B. et al. (2016). Mapping a multiplexed zoo of mRNA expression.Development,143 (19), 3632-3637. doi: 10.1242/dev.140137