Transfer of BAC insert fragments into the BIBAC-S vector using Not I. Two BAC clones with insert sizes of ~162 kb (lane 1) and ~165 kb (lane 8) were randomly selected from a rice variety Minghui 63 BAC library that was constructed using the pIndigoBAC536 vector. DNA samples were digested with Not I and were ligated to the dephosphorylated BIBAC-S vector that was prepared with Not I from the composite BIBAC vector pHZAUBIBAC1. The ligation products were used to transform E. coli DH10B competent cells. DNA samples from thirty randomly selected colonies for each ligation were analyzed using Not I, and the clones that contain BAC insert fragment(s) (with the exception of lane 10) are shown (lanes 2-7 and 9-15, respectively). The molecular weight marker is Midrange I (New England Biolabs).