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
Joel Lopez, Melissa Higgins, Chere Griffin and Vince Prezioso, PhD
BioSystems Laboratory, Brinkmann Instruments, Inc., Westbury, New York
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.
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 membranePage: All 1 2 3 4 Related biology technology :1
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