PCR purification

Materials

“QIAquick gel extraction/PCR purification” – Kit with the PB buffer 1.5 mL labeled Eppendorf tubes

Procedure

• The purification of PCR fragments is obtained with the aid of a kit
• Add 5 volumes of PB buffer to the PCR product and transfer the whole volume to a column
• Centrifugation: 60 seconds, 13000 rpm
• Discard the flowthrough and add 0.75 mL buffer PE to the column and centrifuge again
• Discard the flowthrough and centrifuge again for 60 sec
• The column is added to a sterile, labeled Eppendorf tube.
• Add 30 μl EB-buffer directly to the column, incubate 1 min and elute by centrifugation (60 sec, 13000 rpm).
• Store the eluate on ice