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Spatial transcriptomics of the nematode Caenorhabditis elegans using RNA tomography.

STAR protocols (2021-04-20)
Erik S Schild, Jonas Mars, Annabel Ebbing, Judith Vivié, Marco Betist, Hendrik C Korswagen
ZUSAMMENFASSUNG

RNA tomography or tomo-seq combines mRNA sequencing and cryo-sectioning to spatially resolve gene expression. We have adapted this method for the nematode Caenorhabditis elegans to generate anteroposterior gene expression maps at near-cellular resolution. Here, we provide a detailed overview of the method and present two approaches: one that includes RNA isolation for maximum sensitivity and one that is suitable for partial automatization and is therefore less time-consuming. For complete details on the use and execution of this protocol, please refer to Ebbing et al. (2018).

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Sigma-Aldrich
Ethylendiamintetraessigsäure Dinatriumsalz -Lösung, BioUltra, for molecular biology, pH 8.0, ~0.5 M in H2O
Eppendorf® DNA-LoBind-Röhrchen, capacity 5 mL, cap (screw cap), PCR clean, case of 200 ea (2 bags of 100 Tubes each)