You don't know which insert you have, and the inserts are different sizes, meaning the amount needed for a 1:3 ratio is different. What do you do? Why? (assume there are enough materials for more than one reaction per person).
For ligation process the 1:3 vector to insert ratio is the good to utilize . By considering that we can take 1 ratio of vector and 3 ratio of insert ( consider different insert size ) and take 10 different vials of ligation ( each calculated using different insert size from low to high ) and plot a graph for transformation efficiency and using optimum transformation efficiency we can find out the insert size.
