Team:CSU Fort Collins/Notebook/Protocols=Purify

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Revision as of 16:49, 17 October 2014

PCR Product Purification

PCR Product Purification Protocol

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← Full Table of Contents
Gibson Assembly
Cloning Genes
Plasmid Miniprep
Yeast DNA Extraction
Gel Electrophoresis
PCR Product Purification

    • Adjust total volume of all PCR tubes to 100 μL
    • Add 500 μL Binding Buffer to each tube
    • Mix well
    • Insert one High Pure filter tube into one collection tube
    • Transfer solution from 1 into the filter
    • Centrifuge 60 seconds at maximum speed
    • Disconnect filter tube and discard flow-through
    • Reconnect tubes
    • Add 500 μL wash buffer to upper reservoir
    • Centrifuge 60 seconds at maximum speed
    • Discard the flow-through
    • Recombine tubes
    • Add 200 μL wash buffer
    • Centrifuge for 60 seconds at maximum speed
    • Discard flow-through and collection tube
    • Reconnect filter tube to a clean 1.5 mL centrifuge tube
    • Add 100 μL elution buffer
    • Centrifuge for 1 minute at maximum speed
    • The purified PCR product is now in the microcentrifuge tube
    • Store at 4 °C for short term or at -20 °C for later use


Thank you to Roche (High Pure PCR Product Purification Kit) for this protocol.



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