Protocol for isolation of mouse pre-implantation embryos for gene expression analysis
Summary: Visualizing and quantifying the numerous factors that regulate murine pre-implantation embryonic development is technically challenging. Here, we present a protocol for the isolation of pre-implantation embryos at multiple stages of embryonic development to study gene expression. We describ...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S266616672300446X |
| _version_ | 1797772428043091968 |
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| author | Pallavi Chinnu Varghese Debasree Dutta |
| author_facet | Pallavi Chinnu Varghese Debasree Dutta |
| author_sort | Pallavi Chinnu Varghese |
| collection | DOAJ |
| description | Summary: Visualizing and quantifying the numerous factors that regulate murine pre-implantation embryonic development is technically challenging. Here, we present a protocol for the isolation of pre-implantation embryos at multiple stages of embryonic development to study gene expression. We describe steps for isolating RNA and cDNA synthesis from a small number of embryos. We then detail an immunofluorescence assay for the detection and localization of protein of interest by confocal microscopy in the pre-implantation embryos.For complete details on the use and execution of this protocol, please refer to Varghese et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-12T21:50:42Z |
| format | Article |
| id | doaj.art-f0173158c6a846778d896ecd6652a172 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-12T21:50:42Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-f0173158c6a846778d896ecd6652a1722023-07-26T04:09:38ZengElsevierSTAR Protocols2666-16672023-09-0143102479Protocol for isolation of mouse pre-implantation embryos for gene expression analysisPallavi Chinnu Varghese0Debasree Dutta1Regenerative Biology Program, Rajiv Gandhi Centre for Biotechnology (RGCB), Thiruvananthapuram, Kerala 695014, India; Manipal Academy of Higher Education, Manipal, Karnataka 576104, IndiaRegenerative Biology Program, Rajiv Gandhi Centre for Biotechnology (RGCB), Thiruvananthapuram, Kerala 695014, India; Corresponding authorSummary: Visualizing and quantifying the numerous factors that regulate murine pre-implantation embryonic development is technically challenging. Here, we present a protocol for the isolation of pre-implantation embryos at multiple stages of embryonic development to study gene expression. We describe steps for isolating RNA and cDNA synthesis from a small number of embryos. We then detail an immunofluorescence assay for the detection and localization of protein of interest by confocal microscopy in the pre-implantation embryos.For complete details on the use and execution of this protocol, please refer to Varghese et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S266616672300446XDevelopmental BiologyMicroscopyModel OrganismsMolecular BiologyGene ExpressionAntibody |
| spellingShingle | Pallavi Chinnu Varghese Debasree Dutta Protocol for isolation of mouse pre-implantation embryos for gene expression analysis STAR Protocols Developmental Biology Microscopy Model Organisms Molecular Biology Gene Expression Antibody |
| title | Protocol for isolation of mouse pre-implantation embryos for gene expression analysis |
| title_full | Protocol for isolation of mouse pre-implantation embryos for gene expression analysis |
| title_fullStr | Protocol for isolation of mouse pre-implantation embryos for gene expression analysis |
| title_full_unstemmed | Protocol for isolation of mouse pre-implantation embryos for gene expression analysis |
| title_short | Protocol for isolation of mouse pre-implantation embryos for gene expression analysis |
| title_sort | protocol for isolation of mouse pre implantation embryos for gene expression analysis |
| topic | Developmental Biology Microscopy Model Organisms Molecular Biology Gene Expression Antibody |
| url | http://www.sciencedirect.com/science/article/pii/S266616672300446X |
| work_keys_str_mv | AT pallavichinnuvarghese protocolforisolationofmousepreimplantationembryosforgeneexpressionanalysis AT debasreedutta protocolforisolationofmousepreimplantationembryosforgeneexpressionanalysis |