Endotoxin-Free Miniprep Kit

December 08, 2015

BlueskyReddit
University of North Carolina at Chapel Hill
Research
Laboratory Technician

Overall

Quality of Results

Ease-of-Optimization

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Company:

Zymo Research

Product Name:

ZR Plasmid Miniprep - Classic

Catalog Number:

D4015

In our lab, we perform a good deal of plasmid work on a daily basis. More often than not, we don't need more than a couple of micrograms of plasmid DNA to sequence and then transfect. This kit can yield a decent amount of plasmid DNA from a miniculture of 2-5mL, and it's endotoxin-free so that you can actually transfect cells with the DNA. It's a great way to screen for specific plasmids without having to prep a 200-300mL culture, and it has saved us plenty of time. I don't always get enough DNA to transfect, but there's plenty to sequence.

Experimental Design and Results Summary

Application

Plasmid DNA Isolation

Starting Material

2-5 mL Bacterial Culture

Protocol Overview

This kit is extremely simple, and I can count the steps on my fingers. The first few reagents are multicolored to help visualize cell lysis, and it takes less than 30 minutes total to go from a bacterial culture to endotoxin-free DNA.

Tips

At the endotoxin wash and elution steps, add the liquid to the column and give it a minute or two to incubate before spinning down.

Results Summary

The quality of DNA isn't the best, but I have had no issue whatsoever getting clean sequencing reads from the isolated DNA. For that purpose, it's perfect.

Additional Notes

It's easy to prepare dozens of samples at once if you so prefer.

Image Gallery

Summary

The Good

Quick, simple, and cheap.

The Bad

DNA quality and amount recovered could be better

The Bottom Line

If you want a quick way to prepare endotoxin-free plasmid DNA from small bacterial cultures, this seems to be the way to go. This kit has its limitations, but it works well for that purpose.

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