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Comparison of Growth Techniques and Media for the Purpose of Plasmid ,,, Isolation from E. coli Using the Eppendorf ,,, Perfectprep Plasmid Mini ,,, Kit

Comparison of Growth Techniques and Media for the Purpose of Plasmid Isolation from E. coli Using the Eppendorf Perfectprep Plasmid Mini Kit

Joel Lopez, Melissa Higgins, Chere Griffin and Vince Prezioso, PhD
BioSystems Laboratory, Brinkmann Instruments, Inc., Westbury, New York

Introduction

Isolation and purification of plasmid DNA from bacteria involves three steps: growth of the bacterial culture, harvesting and lysis of the bacteria, and purification of plasmid DNA. Growth of the bacterial cultures is a key factor in obtaining high quality DNA. Using Eppendorfs Thermomixer R in conjunction with Eppendorfs LidBac is an ideal combination.1,2 The LidBac provides excellent gas interchange through a 0.22 m hydrophobic membrane while preventing contamination. The LidBac also allows for growth in standard microcentrifuge tubes, enhanced by excellent heat transmission of the Thermomixer R and suitable Thermoblock.

Objectives

This paper will present an alternative method of growing bacteria in a more reliable manner and at a lower overall cost due to the direct use of the culture vessel in the isolation procedure. The LidBac and Thermomixer R is a dependable combination that provides researchers with an easy-to-use system to grow bacterial cultures directly on the bench top in small quantities.

Materials and Methods

Cell Cultivation and Culture Medias

Escherichia coli (E. coli) strain DH5a containing Plasmid pCS2++ were inoculated into 1.5 ml of LB (Luria-Bertani Medium), TB (Terrific Broth) and 2X YT and cultivated in membrane
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