Protocol to detect RNAs from tissue sections in mice using Y-branched probe in situ hybridization
Summary: Here, we describe a fluorescent in situ hybridization protocol named Yn-situ, standing for Y-branched probe in situ hybridization, to detect RNAs from mice tissue sections. We provide steps for the synthesis and quantification of preamplifier probe using nickase. We also detail the preparat...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722005664 |
| _version_ | 1797980476255764480 |
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| author | Yunming Wu C. Ron Yu |
| author_facet | Yunming Wu C. Ron Yu |
| author_sort | Yunming Wu |
| collection | DOAJ |
| description | Summary: Here, we describe a fluorescent in situ hybridization protocol named Yn-situ, standing for Y-branched probe in situ hybridization, to detect RNAs from mice tissue sections. We provide steps for the synthesis and quantification of preamplifier probe using nickase. We also detail the preparation of tissue section, probe hybridization, signal development using hybridization chain reaction (HCR), and quantification of the signals. This approach avoids the use of proprietary in situ hybridization techniques, therefore reducing costs.For complete details on the use and execution of this protocol, please refer to Wu et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-11T05:54:59Z |
| format | Article |
| id | doaj.art-12777af68ec541d983a2146cf37d7ec1 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-11T05:54:59Z |
| publishDate | 2022-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-12777af68ec541d983a2146cf37d7ec12022-12-22T04:41:54ZengElsevierSTAR Protocols2666-16672022-12-0134101686Protocol to detect RNAs from tissue sections in mice using Y-branched probe in situ hybridizationYunming Wu0C. Ron Yu1Stowers Institute for Medical Research, Kansas City, MO 64110, USA; Corresponding authorStowers Institute for Medical Research, Kansas City, MO 64110, USA; Department of Anatomy and Cell Biology, University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, KS 66160, USA; Corresponding authorSummary: Here, we describe a fluorescent in situ hybridization protocol named Yn-situ, standing for Y-branched probe in situ hybridization, to detect RNAs from mice tissue sections. We provide steps for the synthesis and quantification of preamplifier probe using nickase. We also detail the preparation of tissue section, probe hybridization, signal development using hybridization chain reaction (HCR), and quantification of the signals. This approach avoids the use of proprietary in situ hybridization techniques, therefore reducing costs.For complete details on the use and execution of this protocol, please refer to Wu et al. (2022). : 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/S2666166722005664Developmental biologyMicroscopyMolecular biologyIn situ hybridizationNeuroscienceMolecular/Chemical probes |
| spellingShingle | Yunming Wu C. Ron Yu Protocol to detect RNAs from tissue sections in mice using Y-branched probe in situ hybridization STAR Protocols Developmental biology Microscopy Molecular biology In situ hybridization Neuroscience Molecular/Chemical probes |
| title | Protocol to detect RNAs from tissue sections in mice using Y-branched probe in situ hybridization |
| title_full | Protocol to detect RNAs from tissue sections in mice using Y-branched probe in situ hybridization |
| title_fullStr | Protocol to detect RNAs from tissue sections in mice using Y-branched probe in situ hybridization |
| title_full_unstemmed | Protocol to detect RNAs from tissue sections in mice using Y-branched probe in situ hybridization |
| title_short | Protocol to detect RNAs from tissue sections in mice using Y-branched probe in situ hybridization |
| title_sort | protocol to detect rnas from tissue sections in mice using y branched probe in situ hybridization |
| topic | Developmental biology Microscopy Molecular biology In situ hybridization Neuroscience Molecular/Chemical probes |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722005664 |
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