Amplification and Detection of Transposon Insertion Flanking Sequences Using Fluorescent MuAFLP
Author(s) -
David Edwards,
Jane A. Coghill,
Jacqueline Batley,
Michael J. Holdsworth
Publication year - 2002
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/02325rr04
Subject(s) - transposable element , biology , genetics , tn3 transposon , sleeping beauty transposon system , transposase , genomic dna , amplified fragment length polymorphism , dna , microbiology and biotechnology , genome , gene , population , demography , sociology , genetic diversity
The amplification of transposon insertion flanking sequences is the basis of a variety of techniques used for the detection and characterization of specific transposon insertion events. We have developed a method for the efficient size determination and quantification of amplified genomic sequences that flank Mutator (Mu) transposon insertions in maize. Using this detection method, we have been able to optimize Mu insertion site amplification and to assess amplification from increasingly complex templates representing increasing numbers of Mu-active maize plants. This detection method should be applicable for the characterization of transposon or transgene insertion events in a wide variety of organisms.
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