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===MmeI digestion=== Digestion with MmeI x2 Total 80ul PCR 48ul 10X NEBuffer 4 8ul 16 1mM SAM(fresh) 16ul 32 2U/ul Mme I 8ul 16 1mM SAM: 32mM SAM 1ul + 31ul ddH2O. 37C 2h. Qiaquick column purification. Elute in 50ul EB. 28ul samples are used for endrepair and 22ul are used for ligation directly. [[Image:20090928_after MmeI.jpg]]20090928_after MmeI
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