Team:Calgary/Notebook/Protocols/Process10
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PCR Purification
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The kit we used over the summer was purchased from Qiagen. This kit is used to purify PCR products, as well DNA products in enzymatic reactions, particularly, restriction digests.
- Add 5 volumes of Buffer PB to 1 volume of PCR product and then mix.
- Place a spin column in a 2 mL collection tube.
- Apply the mixture of PB buffer + PCR product through the QIA column and centrifuge for 1 minute.
- Discard the flow-through and place the column back in the tube.
- Wash the product using 0.75 mL buffer PE and then centrifuge for 1 min.
- Discard the flow-through and place the column back into the tube. Centrifuge the column inside the tube to discard the additional fluid in the column.
- Elute the DNA into a clean microcentrifuge tube with buffer EB.
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