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Team:Newcastle/PCR purification
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(New page: =PCR Purification= ==Materials== * 1.5ml microcentrifuge tubes * 2ml collection tube * QIAquick columns * Buffer PB * Buffer EB * Buffer PE * DNA mixture from PCR ==Protocol== # Add 5 ...)
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(New page: =PCR Purification= ==Materials== * 1.5ml microcentrifuge tubes * 2ml collection tube * QIAquick columns * Buffer PB * Buffer EB * Buffer PE * DNA mixture from PCR ==Protocol== # Add 5 ...)
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Revision as of 10:10, 26 August 2010
PCR Purification
Materials
- 1.5ml microcentrifuge tubes
- 2ml collection tube
- QIAquick columns
- Buffer PB
- Buffer EB
- Buffer PE
- DNA mixture from PCR
Protocol
- Add 5 volumes of Buffer PB to 1 volume of PCR product.
- Put a QIAquick spin column into a 2ml collection tube.
- Apply the mixture to the QIAquick column to bind the DNA and centrifuge for 30 to 60 seconds.
- Discard flow-through. Add 0.75 ml of Buffer PE to the spin column to wash and centrifuge for 30 to 60 seconds.
- Discard flow-through and centrifuge for another 1 minute.
- Place QIAquick spin column into a clean 1.5ml microcentrifuge tube.
- Add 50 µl Buffer EB to the center of the membrane of spin column and centrifuge for 1 minute.
- If analysing purified DNA using gel, add 1 volume of Loading Dye to 5 volumes of DNA.
