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I am trying to figure out the details of the homolougous recombination in order to be able to insert a certain piece of DNA in the host genome after double strand break.. Does anybody know how long the homology arms of the repair template must be ? Can the left arm be longer than the right one? I have read several papers which have different sizes 90 bp vs kb.. In my case, i am not so much preoccupied by off target but the insertion should occur also on the desired site possibly at high frequency and as fast as possible. Thank you, Paola

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marked as duplicate by WYSIWYG, Chris, AliceD, The Last Word, Atl LED Mar 13 '15 at 13:59

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As little as 14bp of homology can work. See this reference.

I would recommend at least a length of 90bp but 1kb is probably overkilled. Yes both arms can have different length (staying in the same order of magnitude).

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