Hello,

I am workig with three elements Multisite Gateway Kit to clone a promoter, gene and terminator.

Now I have good results for the promoters and terminator, even I could add attB regions for two of the fours genes I have to clone.

However two of the genes I could not get the attB PCR products. I have tried several times the PCR reaction testing different annealing T. Also I have tested several forward and reverse primers in which I have add more or less base pairs in the region in which the primers bind to the target gene but results always is the same, No attB PCR product appears. I really appreaciate if someone can give advise in how to handle this problem.

Thanks!!!!