Lyse Bacteria by Sonication

Getting Started

You have bacterial cultures and you want to extract something inside of them (e.g., overexpressed protein, plasmid DNA, tRNAs, total lysate, etc.). You’re going to need to lyse this culture. You can use an sonicator to do so, disrupting bacterial walls and membranes using ultrasonic frequencies pumped into the bacterial culture. This protocol shows you how to use a sonicator to mechanically lyse cells.

Materials and Equipment

Prerequisite Protocols

Prepare lysis buffer: Make Protein Purification Buffers and Media.

Prepare pellets of bacteria expression proteins: Grow and Induce Expression Strains.

Protocol

Resources and References

• Papers
• Original PURE paper

Cell-free translation reconstituted with purified components

Nature Biotechnology - Cell-free translation reconstituted with purified components



doi.org


• OnePot PURE

A Simple, Robust, and Low-Cost Method To Produce the PURE Cell-Free System

We demonstrate a simple, robust, and low-cost method for producing the PURE cell-free transcription–translation system. Our OnePot PURE system achieved a protein synthesis yield of 156 μg/mL at a cost of 0.09 USD/μL, leading to a 14-fold improvement in cost normalized protein synthesis yield over existing PURE systems. The one-pot method makes the PURE system easy to generate and allows it to be readily optimized and modified.



doi.org



OnePot PURE Cell-Free System | Text Page

Scientific Article | The defined PURE (protein synthesis using recombinant elements) transcription-translation system provides an appealing chassis for...



dx.doi.org

Credits

Yan Zhang, Zoila Jurado, and Miki Yun (Richard Murray Lab, Caltech)

• Developers
• Murray Lab