hello all
Iam using PfastBac dual vector for cloning two insert . I could successfully ligate insert1 but which i try to ligate the other one i could never get the ligation worked out . I tried the opposite cloning insert 2 first and then again ligating insert 1 , but then insert 1 doesn't ligate . So now i have one vector with insert1 and other with insert2 . I have tried all ratios like 1:1 , 1:2 , 1:4 , 2:2 , 2:4 , 2:1 etc but nothing worked . Has anyone faced similar problems like this . Would be happy to get some help .
Bhargav
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