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Abstract
It has been observed that fragments cut by Nde I type II restriction enzyme have low ligation efficiency. To investigate this observation, λ DNA was digested to produce Nde I fragments. In ligation trials using T4 DNA ligase, Nde I fragments were shown to have lower ligation efficiency than Hind III fragments. The effect of different ligation incubation times on the efficiency was also examined. Although the cohesive ends of Nde I digested fragments would normally ligate in one hour, higher ligation efficiency was observed when Nde I fragments were allowed to ligate for 22 hours or more. In addition, a specific fragment was found to be present in all various lengths of ligation incubation time, which suggests that it did not ligate as effectively as other fragments. It is not clear why this fragment did not ligate as efficiently as others.