I am a bit confused about the ratios conceding vector/insert...
If I check this website: http://www.addgene.o...s/dna-ligation/ , I can see they use 25ng vector and 75ng insert.. This is a ratio of 1/3 (vector/insert).
And yes: this is a normal ratio, however, it seems to make no sense if you use calculators like this one: http://www.insilico..../Lig_Input.html
The amount (in ng) of oligo (insert) that has to be used for cloning is always a lot lower than the amount of vector (in ng).
Lets say: we have 5000 bps of vector, 25 ng and we have an insert of 500 bps... This calculator then tells you to use 7,5 ng insert!!
This is not the 75 ng ...
I do not get this.
Is the addgene website a bit mistaken and taking ng ratios for molar ratios?