Amplification and detection of transposon insertion flanking sequences using fluorescent muAFLP.
The amplification of transposon insertionflanking sequences is the basis of a variety of techniques usedfor 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 thatflank 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 applicablefor the characterization of transposon or transgene insertion events in a wide variety of organisms.
Edwards
D
D
Department of Agricultural Sciences, University of Bristol, Long Ashton, UK. dave.edwards@bbsrc.ac.uk
Coghill
J
J
Batley
J
J
Holdsworth
M
M
Edwards
K J
KJ
D017931:DNA Primers; D004251:DNA Transposable Elements / Q000235:genetics*; D005453:Fluorescence; D016254:Mutagenesis, Insertional / Q000379:methods* / Q000592:standards; D021141:Nucleic Acid Amplification Techniques / Q000379:methods* / Q000592:standards; D010761:Phosphorus Radioisotopes; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D020079:Terminal Repeat Sequences; D019076:Transgenes / Q000235:genetics; D003313:Zea mays / Q000235:genetics*
D017931:DNA Primers; D004251:DNA Transposable Elements; D010761:Phosphorus Radioisotopes