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In addition, Weng et al

It has also been suggested that repair of the 2nd strand takes advantage of the two-fold symmetry of many short-range complexes, such that its ligation is mediated by the second of the two ligase molecules that were present in the initial complex 12,15

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Using more water, like 4mL, will make the solution less concentrated (250mcg per 0.1mL), which can be useful for smaller, more precise dosing protocols

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