Sequencing unpleasantness

So after weeks of trying to get my other cloning reactions to work (two out of five are stalled at the moment), I finally bit the bullet and decided to actually try to sort out my sequencing data from my first two putative clones of the first region of my protein.

After spending two hours trying to think upside down and backwards, I stumbled across something disturbing: for some bizarre reason, they both have a point mutation! Bah!

This means I have to go double check the template I PCR’d the insert out of. I spent four months creating those clones!

I’m going to distract myself from this possible catastrophe by redoubling my efforts to clone the active and inactive catalytic domain into the overexpression vector.