Miniprep Buffer Mixing

The plasmid miniprep (aka. Ethanol Lysis) is the premier method of plasmid purification and will be your bread and butter for working with most microorganisms. You might even want to try purifying larger volumes of plasmid DNA, via a midiprep or maxiprep.

The key difference between these processes is the silica column. I’m going to make a guide on these, which will be here one day – but in the meantime, the important point to understand is that you can purchase generic silica columns for ~0.50¢/each without having to shill out for a full miniprep kit. Keep in mind that columns can be manufactured to capture different sizes of DNA fragment and choosing the wrong type of column will impact your yield.

Once you’ve got the columns, you’re going to need buffers. Openwetware.org have been our primary source for the buffer recipes featured in our guides, but we also could not have succeeded without the mentoring of Dr. Nicholas Coleman.

Click through to the following pages for a demonstration video, protocols and additional resources for mixing each of the miniprep buffers:

1. Buffer P1 (Resuspension Buffer)

2. Buffer P2 (Lysis Buffer)

3. Buffer N3 (Neutralisation Buffer)

4. Buffer PB (Wash Buffer 1)

5. Buffer PE (Wash Buffer 2)

6. Buffer TE, Buffer EB and Water (Elution buffers)