Click here to close Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly. We suggest using a current version of Chrome, FireFox, or Safari.
XB-ART-46513
Nat Protoc 2013 Feb 01;82:310-24. doi: 10.1038/nprot.2013.003.
Show Gene links Show Anatomy links

A single Drosophila embryo extract for the study of mitosis ex vivo.

Telley IA , Gáspár I , Ephrussi A , Surrey T .


???displayArticle.abstract???
Spindle assembly and chromosome segregation rely on a complex interplay of biochemical and mechanical processes. Analysis of this interplay requires precise manipulation of endogenous cellular components and high-resolution visualization. Here we provide a protocol for generating an extract from individual Drosophila syncytial embryos that supports repeated mitotic nuclear divisions with native characteristics. In contrast to the large-scale, metaphase-arrested Xenopus egg extract system, this assay enables the serial generation of extracts from single embryos of a genetically tractable organism, and each extract contains dozens of autonomously dividing nuclei that can be prepared and imaged within 60-90 min after embryo collection. We describe the microscopy setup and micropipette production that facilitate single-embryo manipulation, the preparation of embryos and the steps for making functional extracts that allow time-lapse microscopy of mitotic divisions ex vivo. The assay enables a unique combination of genetic, biochemical, optical and mechanical manipulations of the mitotic machinery.

???displayArticle.pubmedLink??? 23329004
???displayArticle.link??? Nat Protoc
???displayArticle.grants??? [+]


References [+] :
Azuma, Analysis of SUMOylation of topoisomerase IIalpha with Xenopus egg extracts. 2009, Pubmed, Xenbase