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PCR Purification
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==PCR Purification== | ==PCR Purification== | ||
Latest revision as of 00:59, 30 October 2008
PRINCETON IGEM 2008
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PCR Purification
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
