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Rad51-mediated enhancement of DNA ligation in the presence of ADP requires Ca2+. The linear dsDNA with a 3′ four-nucleotide overhang was incubated with Rad51 and E. coli DNA ligase in the presence of ADP, in the reaction mixture containing the indicted amounts of CaCl2 and MgCl2 for 40 min (panel (i)) and 20 min (panel (ii)). We repeated similar experiments twice to confirm the reproducibility of the results.
