Affordable DNA Prep In The Midi Range

August 25, 2017

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Company:

Thermo Scientific

Product Name:

GeneJET Plasmid Midiprep Kit

Catalog Number:

K0482

In my lab we frequently need quantities of DNA in the hundreds-of-microgram range, and using Maxipreps is not affordable. This kit allows you to extract efficiently plasmid DNA from mid sized cultures, with no precipitation step, and a reasonable yield.

Experimental Design and Results Summary

Application

Plasmid DNA prep

Starting Material

E. Coli overnight culture, 25-100 mL

Protocol Overview

Conventional alkali lysis is performed, add endotoxin binding reagent to cleared lysate, add ethanol, then bind to spin column in a benchtop centrifuge. Three washes of the filter, then drying by further spin, then elution, all carried out in a 15mL conical tube included in the kit

Tips

Make sure you clear the lysate by prolonged centrifuging in a floor centrifuge. Might as well divide it into eppendorf tubes and spin at 13000rpm in a bench minifuge.

Results Summary

Yield is very satisfactory, around 500ug per 100ml processed culture, and at a fraction of cost compared to maxiprep columns

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Good yield and low price

The Bad

Many centrifugation steps (6) are required from cleared lysate to DNA

The Bottom Line

Given the cost/prep it's a very convenient option for those labs that frequently deal with the necessity of mid sized plasmid preps. Needs around 30' combined use of tabletop centrifuge but the centrifugation steps are very short.

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