Whole-Mount In Situ Hybridization (WISH) for Salamander Embryos and Larvae

Sruthi Purushothaman; Ashley W. Seifert

doi:10.1007/978-1-0716-2659-7_5

Whole-Mount In Situ Hybridization (WISH) for Salamander Embryos and Larvae

Sruthi Purushothaman
, Ashley W. Seifert

Producción científica: Chapter › revisión exhaustiva

1 Cita (Scopus)

Resumen

Whole-mount in situ hybridization (WISH) is widely used to visualize transcribed gene sequences (mRNA) in developing embryos, larvae, and other nucleotide probe permeable tissue samples. This methodology involves the hybridization of an antisense nucleotide probe to the target mRNA, followed by chromogen or fluorescence-based detection. Here we describe a protocol for the spatiotemporal analysis of mRNA transcripts in axolotl embryos/larvae using digoxigenin-labeled riboprobes, anti-digoxigenin alkaline phosphatase, Fab fragments antibody, and NBT/BCIP chromogen detection.

Idioma original	English
Título de la publicación alojada	Methods in Molecular Biology
Páginas	95-107
Número de páginas	13
DOI	https://doi.org/10.1007/978-1-0716-2659-7_5
Estado	Published - 2023

Serie de la publicación

Nombre	Methods in Molecular Biology
Volumen	2562
ISSN (versión impresa)	1064-3745
ISSN (versión digital)	1940-6029

Nota bibliográfica

Publisher Copyright:
© 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Financiación

This work was partially supported by NIH R01AR070313 to Ashley W. Seifert.

Financiadores	Número del financiador
National Institutes of Health (NIH)	R01AR070313
National Institutes of Health (NIH)

ASJC Scopus subject areas

Molecular Biology
Genetics

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title = "Whole-Mount In Situ Hybridization (WISH) for Salamander Embryos and Larvae",

abstract = "Whole-mount in situ hybridization (WISH) is widely used to visualize transcribed gene sequences (mRNA) in developing embryos, larvae, and other nucleotide probe permeable tissue samples. This methodology involves the hybridization of an antisense nucleotide probe to the target mRNA, followed by chromogen or fluorescence-based detection. Here we describe a protocol for the spatiotemporal analysis of mRNA transcripts in axolotl embryos/larvae using digoxigenin-labeled riboprobes, anti-digoxigenin alkaline phosphatase, Fab fragments antibody, and NBT/BCIP chromogen detection.",

keywords = "Digoxigenin, In situ hybridization (ISH), In vitro transcription, Limb development, Limb regeneration, Riboprobe, Salamander",

author = "Sruthi Purushothaman and Seifert, \{Ashley W.\}",

note = "Publisher Copyright: {\textcopyright} 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",

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AU - Seifert, Ashley W.

PY - 2023

Y1 - 2023

N2 - Whole-mount in situ hybridization (WISH) is widely used to visualize transcribed gene sequences (mRNA) in developing embryos, larvae, and other nucleotide probe permeable tissue samples. This methodology involves the hybridization of an antisense nucleotide probe to the target mRNA, followed by chromogen or fluorescence-based detection. Here we describe a protocol for the spatiotemporal analysis of mRNA transcripts in axolotl embryos/larvae using digoxigenin-labeled riboprobes, anti-digoxigenin alkaline phosphatase, Fab fragments antibody, and NBT/BCIP chromogen detection.

AB - Whole-mount in situ hybridization (WISH) is widely used to visualize transcribed gene sequences (mRNA) in developing embryos, larvae, and other nucleotide probe permeable tissue samples. This methodology involves the hybridization of an antisense nucleotide probe to the target mRNA, followed by chromogen or fluorescence-based detection. Here we describe a protocol for the spatiotemporal analysis of mRNA transcripts in axolotl embryos/larvae using digoxigenin-labeled riboprobes, anti-digoxigenin alkaline phosphatase, Fab fragments antibody, and NBT/BCIP chromogen detection.

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KW - In situ hybridization (ISH)

KW - In vitro transcription

KW - Limb development

KW - Limb regeneration

KW - Riboprobe

KW - Salamander

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T3 - Methods in Molecular Biology

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Whole-Mount In Situ Hybridization (WISH) for Salamander Embryos and Larvae

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Financiación

ASJC Scopus subject areas

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