Breaking Caenorhabditis elegans The Easy Way Using The Balch Homogenizer – An Old Tool For a New Application

摘要

The nematode Caenorhabditis elegans is a model organism best known for its powerful genetics. There is an increasing need in the worm community to couple genetics with biochemistry. Isolation of functionally-active proteins, or nucleic acids without the use of strong, oxidizing denaturants, or of sub-cellular compartments from C. elegans has, however, been challenging because of the worms' thick surrounding cuticle. The Balch Homogenizer is a tool that has found much use in mammalian cell culture biology. The interchangeable, single ball bearing design of this instrument permits rapid permeabilization, or homogenization, of cells. Here we demonstrate the utility of the Balch Homogenizer for studies with C. elegans. We describe procedures for the efficient breakage and homogenization of every larval stage, including dauers, and show that the Balch Homogenizer can be used to extract functionally-active proteins. Enzymatic assays for catalase and dihydrolipoamide dehydrogenase show that sample preparation using the Balch Homogenizer equals or out-performs conventional methods employing boiling, sonication or Dounce homogenization. We also describe phenol-free techniques for isolation of genomic DNA and RNA. Finally, we use the tool to isolate coupled mitochondria and polysomes. The reuseable Balch Homogenizer represents a quick and convenient solution for undertaking biochemical studies on C. elegans.