I have been trying to ligate 400 bp fragment into 10.5 kb vector digested with SalI and dephosphorylated with SAP. But I don't get any colonies. I don't know if Sal I is giving some sort of unexplainable star activity. I have cloned this fragment before in pBluescript after PCR and Sal I digestion. Self ligation before dephosphorylation did work once. Can any one give me suggestions? Thanks.
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