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).

Question

Jazlynn Hammond

Engineering

23 June, 07:54

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).

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Stein · Answer 1

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.