Abstract
Here we describe how to prepare nucleoplasmic extracts from suspension cultures of Drosophila S2 (Schneider Line 2) cells. Harvested cells are washed in phosphate-buffered saline supplemented with MgCl2, resuspended in hypotonic buffer, and homogenized. Nuclei are isolated by centrifugation and then sonicated. The nuclear sonicate is placed on a 30% sucrose cushion and sedimented. The soluble nuclear ribonucleoprotein (RNP) complexes remain in the supernatant and the nuclear membrane fragments and chromatin pellet through the sucrose cushion.
Publisher
Cold Spring Harbor Laboratory
Subject
General Biochemistry, Genetics and Molecular Biology
Cited by
1 articles.
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