How to get rid of primer dimers - (Jun/06/2007 )

Hi there ,

Do you know any method to get rid of primer dimers in PCR purification. Well best is to avoid them getting generated! I know. Since I need to sequence my PCR, these could have a bad effect.... e.g. short runs...

I use QIAquick PCR purification KIT.

You can first run the product on agarose gel and separate the two bands (i.e. primer dimer and amplicon) well enough. Then you can excise the amplicon bands from the gel and then do gel purification so you'll only have your specific product.

Try the NucleoSpin Extract II kit from Takara or Macherey-Nagel. The kit has a dilution series for it's elution buffer that raises the cut off for fragment sizes. Using the correct dilution, your target will bind but the primer/dimer will slide right past the membrane during the binding step.

Cheers,

Steve


Steve Danaher
Research Technician
Johns Hopkins University
CRB II Room 184
1550 Orleans Street
Baltimore, MD 21231
phone: 410-955-9686
sdanahe1@jhmi.edu