Small Scale Plasmid DNA Extraction From Salmonella Enterica Culture

University of Birmingham
Biosciences
Research Assistant

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Company:

Qiagen

Product Name:

QIAprep Spin Miniprep Kit (50)

Catalog Number:

27104

I used Qiagen's QIAprep Spin Miniprep plasmid extraction kit for small scale pBAD DNA extraction from S. enterica cultures grown in LB for cloning experiments. This kit method based extraction gave me an yield of 6 to 7 microgram total with quality of 260:280 ratio 1.87.

Experimental Design and Results Summary

Application

Rapid plasmid DNA extraction from bacterial cultures for cloning experiments

Starting Material

Overnight grown 3 ml S. enterica cultures in LB broth

Protocol Overview

S. enterica cultures grown overnight in the LB broth (3 ml) were pelleted by centrifugation at 10000 rpm at room temperature for 4 minutes. Then these pellets were used in the extraction using miniprep kit's protocol with lysis, neutralisation, washing buffers provided with the kit. Finally, plasmid DNA was eluted in 50 microliter of nuclease free molecular biology grade water.

Tips

If you want to increase the concentration of total plasmid DNA, you elute with smaller volume like 20 or 30 microliter volumes.

Results Summary

Finally I got good yield using this kit. I got around 130 to 140 ng/microliter yield with 260:280 ratio of 1.87 (checked by Nanodrop ND1000 spectrophotometer.

Additional Notes

None

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Summary

The Good

Easy to follow and rapid extraction of small quantity of plasmid DNA

The Bad

None

The Bottom Line

Recommended if one intended to get rapid plasmid DNA from the cultures.

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