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Team:CSU Fort Collins/Notebook/Protocols=Purify
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Revision as of 18:19, 9 October 2014
Plasmid Miniprep Protocol
Show Table of Contents
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- Adjust total volume of all PCR tubes to 100 μL
- Add 500 μL Binding Buffer to each tube
- Mix well
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- Insert one High Pure filter tube into one collection tube
- Transfer solution from 1 into the filter
- Centrifuge 60 seconds at maximum speed
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- Disconnect filter tube and discard flow-through
- Reconnectn tubes
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- Add 500 μL wash buffer to upper reservoir
- Centrifuge 60 seconds at maximum speed
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- Discard the flow-through
- Recombine tubes
- Add 200 μL wash buffer
- Centrifuge for 60 seconds at maximum speed
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- Discard flow-through and collection tube
- Reconnect filter tube to a clean 1.5 mL centrifuge tube
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- Add 100 μL elution buffer
- Centrifuge for 1 minute at maximum speed
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- The purified PCR product is now in the microcentrifuge tube
- Store at 4 °C for short term or at -20 °C for later use
