Localizing Ac insertions is a fundamental task in studying Ac-induced mutation and chromosomal rearrangements involving Ac elements. Researchers may sometimes be faced with the situation in which the sequence flanking one side of an Ac/Ds element is known, but the other flank is unknown. Or, a researcher may have a small sequence surrounding the Ac/Ds insertion site and needs to obtain additional flanking genomic sequences. One way to rapidly clone unknown Ac/Ds flanking sequences is via a PCR-based method termed Ac casting. This approach utilizes the somatic transposition activity of Ac during plant development, and provides an efficient means for short-range genome walking. Here we describe the principle of Ac casting, and show how it can be applied to isolate Ac macrotransposon insertion sites. © Springer Science+Business Media, New York 2013.
CITATION STYLE
Wang, D., & Peterson, T. (2013). Isolation of sequences flanking Ac insertion sites by Ac casting. Methods in Molecular Biology, 1057, 117–122. https://doi.org/10.1007/978-1-62703-568-2_8
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