I was wondering if it is possible to know the best vector:insert ratio to use in a ligation reaction, knowing the sizes of both fragments.
Is there any way to calculate it, or you just have to try different ratios until you get the right one?
well, i’m working with a 3,7kb vector and a 1,6kb insert and 1:1 and 1:3 don’t seem to work. I’ve now tried 1:5 and this seems to work but wih very low efficiency.
I’m trying to ligate blunt ends… maybe that’s why the ratios have to be higher..
It would be very helpful if it was possible to calculate the optimal ratio before starting this kind of work.