During setting up assembly cloning reaction, we need to add the insert and vector in ratio 1:4, which is mentioned in the protocol. But I don't know what is the reasons behind this ratio. I will be glad if someone put some light on this very simple question of mine and sends me some link regarding the same.
It is customary to add an excess of insert in order to maximize the chance that the vector will be ligated to the insert, thus generating the desired recombinant plasmid. However, a too high concentration of insert will generate concatemers. Even though these will be mostly resolved by recombination once inside the host, they will decrease the efficiency of transformation. Thus a 3-5 fold excess of insert is usually recommended, but I don't know of any systematic study to determine whether this is really optimum, as it may possibly depend on factors such as insert or vector size, or blunt vs sticky end ligation.
I can forward you to this already existing research gate thread, which I think might answer your question:
https://www.researchgate.net/post/How_to_decide_the_molar_ratio_for_vectorinsert_for_ligation_reaction
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