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Team:Northwestern/Notebook/Protocols/PCR Purification
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We used [http://www.qiagen.com/Products/DnaCleanup/GelPcrSiCleanupSystems/QIAquickPCRPurificationKit.aspx?r=926 QIAquick PCR Purification Kit] from Qiagen. This can be used to purify PCR products or restriction digests. | We used [http://www.qiagen.com/Products/DnaCleanup/GelPcrSiCleanupSystems/QIAquickPCRPurificationKit.aspx?r=926 QIAquick PCR Purification Kit] from Qiagen. This can be used to purify PCR products or restriction digests. | ||
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#Add 5 volumes of Buffer PB to 1 volume of PCR sample and mix. Apply sample to a spin column and centrifuge for 30-60 seconds. Discard flow-through. Place the column back into the same tube. | #Add 5 volumes of Buffer PB to 1 volume of PCR sample and mix. Apply sample to a spin column and centrifuge for 30-60 seconds. Discard flow-through. Place the column back into the same tube. | ||
#Wash with 0.75 mL Buffer PE and centrifuge for 30-60 seconds. Discard flow-through and place the column back into the same tube. Dry spin for an additional 1 minute at maximum speed. | #Wash with 0.75 mL Buffer PE and centrifuge for 30-60 seconds. Discard flow-through and place the column back into the same tube. Dry spin for an additional 1 minute at maximum speed. | ||
#Place column in a clean 1.5 mL microcentrifuge tube. Add 50 uL of nuclease free water to the center of the membrane. Let sit at room temperature for 10 minutes, then centrifuge teh column for one minute to elute. | #Place column in a clean 1.5 mL microcentrifuge tube. Add 50 uL of nuclease free water to the center of the membrane. Let sit at room temperature for 10 minutes, then centrifuge teh column for one minute to elute. | ||
Latest revision as of 19:38, 23 August 2011
PROJECT
RESULTS
CONSIDERATIONS
ABOUT US
NOTEBOOK
ATTRIBUTIONS
PCR Purification
We used [http://www.qiagen.com/Products/DnaCleanup/GelPcrSiCleanupSystems/QIAquickPCRPurificationKit.aspx?r=926 QIAquick PCR Purification Kit] from Qiagen. This can be used to purify PCR products or restriction digests.
- Add 5 volumes of Buffer PB to 1 volume of PCR sample and mix. Apply sample to a spin column and centrifuge for 30-60 seconds. Discard flow-through. Place the column back into the same tube.
- Wash with 0.75 mL Buffer PE and centrifuge for 30-60 seconds. Discard flow-through and place the column back into the same tube. Dry spin for an additional 1 minute at maximum speed.
- Place column in a clean 1.5 mL microcentrifuge tube. Add 50 uL of nuclease free water to the center of the membrane. Let sit at room temperature for 10 minutes, then centrifuge teh column for one minute to elute.
