Photo-isolation chemistry for high-resolution and deep spatial transcriptome with mouse tissue sections.

IF: 0

Abstract

Photo-isolation chemistry (PIC) enables isolation of transcriptome information from locally defined areas by photo-irradiation. Here, we present an optimized PIC protocol for formalin-fixed frozen and paraffin mouse sections and fresh-frozen mouse sections. We describe tissue section preparation and permeabilization, followed by in situ reverse transcription using photo-caged primers. We then detail immunostaining and UV-mediated uncaging to the target areas, followed by linear amplification of uncaged cDNAs, library preparation, and quantification. This protocol can be applied to various animal tissue types. For complete details on the use and execution of this protocol, please refer to Honda et al. (2021).

Keywords

Gene Expression

Molecular Biology

Molecular/Chemical Probes

RNAseq

Sequence analysis

Sequencing

MeSH terms

Animals

DNA, Complementary

Formaldehyde

Frozen Sections

Gene Library

Mice

Transcriptome

Authors

Honda, Mizuki

Kimura, Ryuichi

Harada, Akihito

Maehara, Kazumitsu

Tanaka, Kaori

Ohkawa, Yasuyuki

Oki, Shinya

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1. Photo-isolation chemistry for high-resolution and deep spatial transcriptome with tissue sections