Team:Waterloo/Notebook/Protocols/DNA Digestion

 Digestion of Plasmid DNA 

Before you start  - Know what is the total digestion volume mix. From that you can determine the enzyme concentration which will have to be at or below 10%. You can also calculate the buffer volume based on the concentration of the buffer needed

Materials  MQ water (Vtotal - Vbuffer - VDNA - Venzymes) Buffer (depends on required concentration: For 10x buffer, add 1/10 Vtotal DNA (1 µL) Enzymes (0.5 µL each/digest) Microfuge tube (1/digest)

Instructions  1. In one tube, mix the above materials in the given order. - Keep the enzymes cold as much as possible. Do not leave them out for longer than necessary. 2. Vortex briefly. 3. Centrifuge 2-3 seconds. 4. Digest 1 hour at 37ºC.