Large-Scale Plasmid Isolation from Bacterial Cells

University of Birmingham
School of Biosciences
Postdoctoral Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Thermo Fisher Scientific

Product Name:

GeneJET Plasmid Maixprep Kit

Catalog Number:

K0491

I used GeneJET Plasmid Maxiprep Kit from Thermo Scientific for large scale high quality plasmid especially low copy number from recombinant E. coli cells. The yield was great with 100 microgram per 100 ml overnight culture. The quality was 1.83 for 260:280 ratio in nanodrop assessment.

Experimental Design and Results Summary

Application

Large scale high quality plasmid DNA extraction from gram negative bacteria

Starting Material

100 ml of overnight culture of E. coli cells containing low copy plasmid vector

Tips

Use culture volume according to OD600 calculation table mentioned in the kit's protocol. This is to prevent over saturation of DNA columns.

Results Summary

Plasmid extraction was very simple and easy to follow as mentioned in the kit's protcol. It is also having low speed centrifugation step to reduce the time during extraction protocol as compared to gravity based kits. Yiled was very good with total of 100 micrgram in a 500 microliter nuclease free water. The quality i.e 260:280 ratio was 1.83 and 260:230 ratio was 1.98.

DOI or PMID #

N/A

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Summary

The Good

Easy to follow protcol. seperation can be enhanced by low speed centrifugation.

The Bad

None

The Bottom Line

I found this kit does great job for isolation large scale, high quality plasmid DNA especially of low copy number.

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