Lots of DNA in Minimum Time and Effort

Overall

Quality of Results

Ease-of-Optimization

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Colorado State University
Department of Biology
Graduate student

Company:

Qiagen

Product Name:

Qiagen Plasmid Midi Kit (25)

Catalog Number:

12143

Working in the field of basic biology, everyone needs to use plasmids for cloning followed by transfections. I use this kit to purify plasmids from bacterial culture for use in down-stream applications.

Experimental Design and Results Summary

Application

Preparation of plasmids for transfections

Starting Material

E- Coli DH5-alpha cultures transformed with plasmids

Protocol Overview

I had cloned a gene of interest with point mutation in pGL3Basic plasmid and needed to purify the plasmids for transfection in HeLa cells.

Tips

Read the protocol from beginning to end before starting the purification because it is very important. And keep buffers in 4 degree beforehand.

Results Summary

I purified the plasmids in about 3-4 hrs using the protocol. The yield was very good and the purity of the plasmid was well within acceptable limits (260/280 ratio of 1.80-1.90 and the agarose gel)

Additional Notes

None

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Summary

The Good

All required buffers in one kit (user doesn't need to provide anything except isoproanol) and rapid procedure. Having the cartridges in the kit simplifies the process even further.

The Bad

The names of the buffer are a little confusing (would have been better had they named them as wash buffer, equilibration buffer and so forth). Having said that it is not very difficult to remember these names if one is using the kit frequently. And it seems a little expensive.

The Bottom Line

A standard and most used kit for plasmid purification. Gives good yield with best quality of plasmids. I would recommend the use of this kit for any plasmid purification.

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