Hi,

I am doing three different Multiplex PCR, each with five pairs of primers amplifying regions of different genes. I tested the primers individually first and they all work fine. When i put them together for the Multiplex, one of the three primer mixes has the bands smear when I run the product on the gel (I am using 2% agarose gel in TBE1X, 150V for 1h). I have tried different primers concentration and also tested two different enzymes but the result is still the same. Do you have any suggestions on how to improve the multiplex?

Thanks.