PCR Purification

PRINCETON IGEM 2008

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PCR Purification allows for a buffer exchange and the ability to get rid of unwanted enzymes still surrounding the plasmid DNA.

Using QIAquick kit

1. Add 5 volumes of Buffer PBI to 1 volume of sample.

2. Pipette into a QIAquick spin column(max 770 µl) and centrifuge for 60 sec at 10,000g

3. Discard flow-through.

4. Wash: add 0.75ml Buffer PE(make sure that the buffer has ethanol added to it) to column and centrifuge for 1 min

5. Discard flow-through & centrifuge for 1 min

6. Place column into clean Eppendorf tube

7. Add 50ul Buffer EB or water to center of membrane

8. Let stand at RT for 3 min

9. Centrifuge for 5 min.

Measure the concentration using the UV spectrophotometer