QIAprep Spin Miniprep Kit From Qiagen

The QIAprep Miniprep system is intended for isolation of plasmids from bacteria cells. The kit uses silica-gel-membrane technology to bind DNA, from which the DNA can be eluted with an elution buffer or water. Plasmid DNA purified by this system is immediately ready for use. There are at least three different products available, based on throughput requirements: The low throughput system – QIAprep Spin Miniprep Kit, the medium throughput system –QIAprep 8 Miniprep Kit, and the high throughput system –QIAprep 96 turbo Miniprep kit.

We have experience using the QIAprep Spin Miniprep Kit, which can be used to process 1-20 samples in about 30-45 minutes. We have had good success with this kit – the instructions are very easy to follow and all the necessary reagents are supplied (buffer P1 must be kept at 4ºC because the RNase can degrade). The only special equipment required is a small tabletop centrifuge or vacuum manifold.

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The principle of this kit is based on alkaline lysis of bacteria cells followed by adsorption of DNA onto a silica column, and then washing and elution of plasmid DNA. To begin, simply grow up your plasmid of interest in bacteria with the appropriate selection drug, and then follow the procedure. First, the cells should be spun down and resuspended in an alkaline buffer with an RNase for cell lysis. Next, the lysate in alkaline solution is neutralized with a neutralization buffer. Afterwards, the solution is spun down to remove cellular debris. Only the cell lysate is transferred to the spin column, where only DNA will be adsorbed, while RNA, proteins, and other cellular material will remain in the flow-through. The spin column is then washed once with ethanol, and finally, the DNA is eluted into the elution buffer or water with a pH between 7.0 and 8.5. The plasmid can then be visualized on an agarose gel.

This kit is easy to use, fast, and relatively affordable. The kits can be purchased in quantities of 50 – 250 columns. We have had great success with this kit and would highly recommend it.