Griffitts:PCR clean-up: Difference between revisions
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==Procedure== | ==Procedure== | ||
* Label columns and microcentrifuge tubes with the names of the appropriate PCR reactions | * Label columns and microcentrifuge tubes with the names of the appropriate PCR reactions | ||
* Using P-200, add 200 μL buffer PB to each | * Using P-200, add 200 μL buffer PB to each 40-μL PCR and mix well by gently pipeting up and down | ||
* Transfer mixture to a QiaPrep spin column | * Transfer mixture to a QiaPrep spin column | ||
* Centrifuge at full speed for 30 seconds | * Centrifuge at full speed for 30 seconds | ||
Revision as of 07:21, 10 March 2009
Materials
- Buffer PB
- Buffer PE
- TE buffer
- QiaPrep spin column (1 per PCR reaction)
- 1.5 mL microcentrifuge tube (1 per PCR reaction)
Procedure
- Label columns and microcentrifuge tubes with the names of the appropriate PCR reactions
- Using P-200, add 200 μL buffer PB to each 40-μL PCR and mix well by gently pipeting up and down
- Transfer mixture to a QiaPrep spin column
- Centrifuge at full speed for 30 seconds
- Dump and tap
- Add 600 μL buffer PE to each column
- Centrifuge at full speed for 30 seconds
- Dump and tap
- Centrifuge for an additional 30 seconds at full speed
- Move column to a 1.5 mL microfuge tube
- Add 50 μL TE buffer (to elute)
- Drip this directly onto the filter disk but don't touch it with the pipette tip
- Wait 1 minute
- Centrifuge at full speed for 1 minute
- Point the lids of the microcentrifuge tubes in a clockwise direction, otherwise they'll break off
