BEST Bacterial Plasmid Miniprep Kit

June 16, 2016

Boston University
Biomedical Engineering
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Zymo Research

Product Name:

ZR Plasmid Miniprep Classic

Catalog Number:

D4016

I do A LOT of molecular cloning for synthetic biology experiments in mammalian cells. I routinely perform 40-80 minipreps a day. This kit always gives me high yields (500-1200 ng DNA per uL) even when I elute in ddH20 rather than Elution Buffer. It is also more cost efficient than other more popular kits like Qiagen.

Experimental Design and Results Summary

Application

Bacterial Plasmid DNA Miniprep

Starting Material

Expanded bacterial cell culture where cells have been transformed with antibiotic resistant bacterial plasmid

Protocol Overview

Grow 5 mL overnight bacterial culture (I use Stabl3 cells), spin down cells, resuspend pellet in buffer P1, add lysis buffer 2 min, mix in neutralization buffer. Transfer cell lysates to microcentrifuge tubes, spin down debris. Transfer supernatant to microcentrigue purification columns, spin, wash buffer, spin, wash buffer 2, spin, elute.

Tips

It helps to spin empty columns after second wash buffer to get rid of all ethanol before eluting. I usually elute in 25 uL EB or ddH20

Results Summary

I always achieve high yields (with high copy plasmids): 500-1200 ng/ uL., typically close to 1000 ng/uL.

Additional Notes

The kit/protocol can be adapted to a vacuum manifold which goes a little faster for small numbers of preps

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Summary

The Good

Excellent DNA yield

The Bad

They don't provide enough waste tubes for the number of columns they give you, I end up using other microcentrifuge tubes towards the end of the kit.

The Bottom Line

This kit is fast, easy, gives excellent robustly high yields of bacterial plasmid DNA and is cheaper than other comparable kits.

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