Immunoprecipitation: Lysing Bacteria by Sonication

Ed Harlow; David Lane

doi:10.1101/pdb.prot4532

Protocol

Immunoprecipitation: Lysing Bacteria by Sonication

Ed Harlow and
David Lane

This protocol was adapted from “Immunoprecipitation,” Chapter 7, in Using Antibodies by Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1999.

INTRODUCTION

In this protocol, bacterial cells are lysed by being subjected to short, intense treatments with ultrasound, which breaks the cell walls and shears the DNA into sizes that will not affect the viscosity of the samples. Note that this method causes some denaturation of the samples. The resulting lysate is ready for preclearing.