The PCR Kleen Spin purification module uses size exclusion chromatography to remove unincorporated primers, nucleotides, salts, and enzymes from PCR products. This protocol purifies PCR products using simple spin columns that remove the contaminating molecules by absorbing them into porous beads. Larger molecules, such as PCR products greater than 200 bp, cannot enter the beads and are eluted from the column into a collection tube.
Following the purification step, both the unpurified and the purified PCR products can be resolved side-by-side using agarose gel electrophoresis and stained to visually assess the removal of oligonucleotide primers. Once purified, the PCR product is ligated into the pJet1.2 plasmid vector.
The PCR Kleen Spin purification module is an integral component of the Cloning and Sequencing Explorer Series, where it is used to purify a portion of GAPDH gene that has been amplified. The module can also be purchased separately and used in other experiments to:
- clean any PCR reaction prior to ligation
- remove primers prior to subsequent nested PCR reactions
- remove nucleotides before direct DNA sequencing