Document Detail


Centrifugal elutriation of large fragile cells: isolation of RNA from fixed embryonic blastomeres.
MedLine Citation:
PMID:  1381875     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
A Nasir; S D Reynolds; P C Keng; L M Angerer; R C Angerer
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Analytical biochemistry     Volume:  203     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1992 May 
Date Detail:
Created Date:  1992-10-13     Completed Date:  1992-10-13     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  22-6     Citation Subset:  IM    
Affiliation:
Department of Biology, University of Rochester, New York 14627.
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MeSH Terms
Descriptor/Qualifier:
Animals
Blastomeres / chemistry*
Cell Separation / methods*
Centrifugation*
Ethanol
Fixatives
RNA / isolation & purification*
Sea Urchins
Grant Support
ID/Acronym/Agency:
GM07102/GM/NIGMS NIH HHS; GM25553/GM/NIGMS NIH HHS; HD00601/HD/NICHD NIH HHS
Chemical
Reg. No./Substance:
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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