Plasmid DNA Extraction from E. coli

March 31, 2016

BlueskyReddit
University of Birmingham
School of Biosciences
Postdoctoral Research Fellow

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Company:

Qiagen

Product Name:

QIAGEN Plasmid Midi Kit (25)

Catalog Number:

12843

I have extracted high copy plasmid from E. coli grown in LB broth overnight using Qiagen Plasmid midi kit with gravity flow based anion exchange column based extraction. 25 ml culture was used and got 27 micrograms of plasmid DNA with very good quality (260: 280 ratio of 1.84.)

Experimental Design and Results Summary

Application

High copy plasmid DNA extraction for cloning

Starting Material

25 ml of overnight grown E. coli culture

Protocol Overview

25 ml of E. coli culture grown overnight in LB broth was pelleted and then used for extraction following easy-to-follow instructions from the kit. Finally, I eluted in 100 microliter of molecular biology grade nuclease free water.

Tips

Pre-heat nuclease free water to 55 degree Celsius

Results Summary

Nano-Drop quantification results after the extraction was 272 ng/microliter with total yield for 27 microgram.

Additional Notes

None

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Summary

The Good

Simple and fast protocol and in less than 3 hrs you will be having your plasmid DNA extracted.

The Bad

None

The Bottom Line

We routinely use this kit in our laboratory as we do large scale cloning methods in bacteria. I highly recommend this kit if you are routinely doing cloning experiments.

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