That is, lack of successfull transformation. I got my colonies on the LB agar, and was happy, everything going on as planned. But, darn, something wasn’t right. Using restriction enzymes to cut the new plasmids, checking if my genes actually had been incorporated, showed … nothing. I’ve spent the last days trying to find out what went wrong. My PCR products seems all right. The plasmid we’re using for transformation has a “suicide” gene which gets transcribed if there is no successful gene incorporation, inhibiting growth. But, I got my colonies, the bacteria did grow. I even tried several different restriction enzymes, and restriction buffers, just to be sure. But, nothing, nada, my gene is not there. Weirdness. I will probably end up doing the transformation from scratch. Albeit I really want to know what went wrong…