| Centrifugal elutriation of large fragile cells: isolation of RNA from fixed embryonic blastomeres. | |
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MedLine Citation:
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PMID: 1381875 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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In order to analyze the RNA populations present in different cells of very early embryos, we have developed a protocol to purify these large blastomeres using counterflow centrifugal elutriation (CCE). This procedure employs ethanol fixation to stabilize the cells against shear forces encountered during CCE. Using this method, we fractionated the three different blastomere types of the 16-cell sea urchin embryo, the micromeres, mesomeres, and macromeres, achieving 96, 94, and 96% mean purities, respectively. We show here that intact RNA is recovered with equal efficiency from each blastomere preparation. Using this method, we have identified several RNAs that are distributed non-uniformly among these cells. |
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Authors:
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A Nasir; S D Reynolds; P C Keng; L M Angerer; R C Angerer |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Analytical biochemistry Volume: 203 ISSN: 0003-2697 ISO Abbreviation: Anal. Biochem. Publication Date: 1992 May |
Date Detail:
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Created Date: 1992-10-13 Completed Date: 1992-10-13 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 0370535 Medline TA: Anal Biochem Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 22-6 Citation Subset: IM |
Affiliation:
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Department of Biology, University of Rochester, New York 14627. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Blastomeres / chemistry* Cell Separation / methods* Centrifugation* Ethanol Fixatives RNA / isolation & purification* Sea Urchins |
| Grant Support | |
ID/Acronym/Agency:
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GM07102/GM/NIGMS NIH HHS; GM25553/GM/NIGMS NIH HHS; HD00601/HD/NICHD NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Fixatives; 63231-63-0/RNA; 64-17-5/Ethanol |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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