ligation question - (Oct/10/2005 )

a simple question. do you measure the concentration of the insert and vector and then use the 3:1 to do the ligation?
I just usually look at the agarose gel and try a different ratio of insert and vector to do the ligation, never really measure the con of each.

just wonder how did you do it? did you find measuing the con easiler to get it working?

Thanks a lot.
cathy

I do as you do...

hi
i use 1:5 ratio. But i estimate the conc by agarose gel.
fred

hi...
i eye estimate the conc. of the insert of the vector and the insert on the agarose gel and set up an ligation of 1:5 and 1: 10 ratio......... usually i have observed that 1:10 works well to me.
raky

technically this is supposed to be a molar ratio, thus the length of the DNAs should be taken into account. In reality there is quite a bit of felxibility in the ratio you use as long as you have more insert to vector.

thanks fella