High-quality frozen extracts of Xenopus laevis eggs reveal size-dependent control of metaphase spindle micromechanics.

ABSTRACT: Cell-free extracts from unfertilized Xenopus laevis eggs offer the opportunity for a variety of biochemical and biophysical assays for analyzing essential cell cycle events such as metaphase spindle assembly. However, the extracts often exhibit substantial variation in quality and have low storage stability, factors that hamper their experimental utility. Here we report a simple two-step method for preparing frozen egg extracts that retain spindle assembly activity levels similar to those of freshly prepared extracts. Extract degradation associated with the freeze-thaw process can be substantially reduced by using centrifugal filter-based dehydration and slow sample cooling. Large amounts of frozen extract stocks from single-batch preparations allowed us to collect extensive data in

SUBMITTER: Takagi J 

PROVIDER: S-EPMC5531733 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature