Griffitts:PCR clean-up

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