Chapter 1 - Mitotic Spindle Assembly on Chromatin Patterns Made with Deep UV Photochemistry

We provide a detailed method to generate arrays of mitotic spindles in vitro. Spindles are formed in extract prepared from unfertilized Xenopus laevis eggs, which contain all the molecular ingredients of mitotic spindles. The method is based on using deep UV photochemistry to attach chromatin-coated...

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Published inMethods in cell biology Vol. 120; pp. 3 - 17
Main Authors Tarnawska, Katarzyna, Pugieux, Celine, Nedelec, Francois
Format Journal Article
LanguageEnglish
Published 01.01.2014
Subjects
Xenopus laevis
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Summary:We provide a detailed method to generate arrays of mitotic spindles in vitro. Spindles are formed in extract prepared from unfertilized Xenopus laevis eggs, which contain all the molecular ingredients of mitotic spindles. The method is based on using deep UV photochemistry to attach chromatin-coated beads on a glass surface according to a pattern of interest. The immobilized beads act as artificial chromosomes, and induce the formation of mitotic spindles in their immediate vicinity. To perform the experiment, a chamber is assembled over the chromatin pattern, Xenopus egg extract is flowed in and after incubation the spindles are imaged with a confocal microscope.
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ISSN:0091-679X
DOI:10.1016/B978-0-12-417136-7.00001-X