Targeted Analysis of Chromatin Events (TACE)
Visualization of meiotic chromatin from pollen mother cells has become an essential technique to study meiosis in the model plant Arabidopsis thaliana. Here we present an advanced cytogenetic method that combines improved immunocytology with chromosome painting, thereby generating a tool to quantitatively analyze localization of proteins to any given genomic region. Proteins involved in different processes such as DNA double-strand break formation and recombinational repair can be visualized on meiotic chromatin with the additional feature of assessing their abundance at specific chromosomal locations.
Key wordsArabidopsis thaliana Chromosome painting FISH Immunocytology Meiosis
We thank the European Union (FP7-ITN 560 606956), the University of Vienna (I031-B), and the Austrian Science Fund (SFB F3408; P18036) for funding (J.S., P.S., and M.T.K.). C.C. is supported by the National Science Foundation (IOS:1025881 and IOS:1546792) and the Grant-in-Aid program at the University of Minnesota. We furthermore thank Chris Franklin for providing the ASY1 antibody and Sue Armstrong, James Higgins, and Katja Schneider for very helpful technical demonstrations and discussions.
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