PCR clean-up (Qiagen)

Materials

• Buffer PB (in Qiagen kit)
• Buffer PE (in Qiagen kit)
• Buffer EB (in Qiagen kit)
• QiaPrep spin column (1 per PCR reaction)
• 1.5 mL microcentrifuge tube (1 per PCR reaction)

Procedure

• Label spin columns and microcentrifuge tubes with the names of the appropriate PCR reactions
• Using a P-200, add 200 μL buffer PB to each 40-μL PCR and mix well by gently pipetting up and down
• Transfer the mixture to a QiaPrep spin column
• Centrifuge at full speed for 30 seconds
• Dump and tap
• Add 700 μL buffer PE to each column
• Centrifuge at full speed for 30 seconds
• Dump and tap
• Centrifuge for an additional 5 minutes at full speed
• Move the column to a 1.5 mL microfuge tube
• Add 50 μL buffer EB (to elute)
  
  • Drip this directly onto the filter disk but don't touch it with the pipette tip
• Wait 2 minutes
• Centrifuge at full speed for 1 minute
  
  • Point the lids of the microcentrifuge tubes in a clockwise direction, otherwise they'll break off