RNA extraction from organoids encapsulated in alginate norbornene using TRIzol Reagent v1

This protocol describes the isolation of high-quality RNA from organoids encapsulated in alginate norbornene hydrogels. Organoids are first released from the hydrogel matrix using alginate lyase digestion, followed by homogenization in TRIzol and RNA extraction through standard organic phase separation. The expected outcome is the recovery of intact RNA with sufficient yield and purity for downstream molecular biology applications, including qPCR and sequencing. Major optimizations include removal of the hydrogel prior to RNA extraction to minimize contamination from residual matrix components, refinement of the homogenization step to improve reproducibility, and the inclusion of glycogen blue to facilitate visualization and recovery of small RNA pellets, particularly in low-yield samples.

Authors

Publication Details

Published
2025-03-06
DOI
https://doi.org/10.17504/protocols.io.dm6gp94w8vzp/v1
Citations
1
Primary Topic
Educational Robotics and Engineering
Type
article
Field-Weighted Citation Impact
2.48

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article

RNA extraction from organoids encapsulated in alginate norbornene using TRIzol Reagent v1

Lorenzo Moroni, Carlos Mota, Helen Kearney
1 citations
Educational Robotics and Engineering
2.48
article

RNA extraction from organoids encapsulated in alginate norbornene using TRIzol Reagent v1

Lorenzo Moroni, Carlos Mota, Helen Kearney
article en
1 citations

Abstract

This protocol describes the isolation of high-quality RNA from organoids encapsulated in alginate norbornene hydrogels. Organoids are first released from the hydrogel matrix using alginate lyase digestion, followed by homogenization in TRIzol and RNA extraction through standard organic phase separation. The expected outcome is the recovery of intact RNA with sufficient yield and purity for downstream molecular biology applications, including qPCR and sequencing. Major optimizations include removal of the hydrogel prior to RNA extraction to minimize contamination from residual matrix components, refinement of the homogenization step to improve reproducibility, and the inclusion of glycogen blue to facilitate visualization and recovery of small RNA pellets, particularly in low-yield samples.

European Commission, H2020 Marie Skłodowska-Curie Actions
Openalex Percentile: Top 8%
Educational Robotics and Engineering
2.48
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