Don't worry, leave it for now, move on to the next step. Then come back to it when your mind is fresh. Often looking at the same thing over and over again you start going blind towards it. Get a senior to read your protocol. Better still, your library should have a copy thesis's for previous students who may have done the same protocol! Most PhD's vary only slightly within a department. Best of Luck!

I always have chocolate if you want it. Always. And i'll make a nice pot of coffee too.

Neil, did u get the ligating thing to work? Was just thinking, instead of your colleagues having a go, they should have tried to be more helpful by helping you figure out what was going wrong. No cake for them!

DanB, have you got any of that 70% cocoa chocolate?

I normally get Green and Blacks, that's quite high cocoa content. I have the white one, the mint one that has fondanty centre and the spiced orange one (which my friend says tastes of christmas!).

Hi H. The ligation hasn't worked yet. But I have worked out what the problem, and will be doing the ligation today, so by the end of the week I should know if it has worked.
Sam, I would love to move onto the next step. The trouble is I have done everything else, and now the rest of my PhD hinges on me being able to finish the small section.