Document Detail

Using multiplex single-base extension typing to screen for mutants defective in RNA editing.
MedLine Citation:
PMID:  23037308     Owner:  NLM     Status:  MEDLINE    
RNA editing is an RNA maturation process that changes the nucleotide present at particular positions (editing sites) in specific RNAs; in plant organelles, the most common nucleotide change is from cytidine (C) to uridine (U). In a mutant suspected of affecting RNA editing, all known editing sites have to be analyzed. Therefore, to screen a population of mutants, all individuals must be analyzed at every editing site. We describe a multiplex single-nucleotide polymorphism (SNP)-typing procedure to economically screen a mutant individual or population for differences at hundreds of nucleotide positions in RNA or DNA. By using this protocol, we have previously identified mutants defective in RNA editing in a randomly mutated population of Arabidopsis thaliana. The procedure requires 2-3 weeks to identify the individual plant in the mutant population. The time required to locate the mutated gene is between 3 and 24 months in Arabidopsis. Although this procedure has been developed to study RNA editing in plants, it could also be used to investigate other RNA modification processes. It could also be adapted to investigate RNA editing in other organisms.
Mizuki Takenaka; Axel Brennicke
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-10-04
Journal Detail:
Title:  Nature protocols     Volume:  7     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-10-05     Completed Date:  2013-03-19     Revised Date:  2013-03-21    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  1931-45     Citation Subset:  IM    
Molekulare Botanik, Universität Ulm, Ulm, Germany.
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MeSH Terms
Arabidopsis / genetics*
DNA Mutational Analysis / methods*
DNA Primers
Genes, Plant
Polymerase Chain Reaction
Polymorphism, Single Nucleotide
RNA Editing / genetics*
Reg. No./Substance:
0/DNA Primers

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