u/InviteDry4161

I have been trying to ligate ( for the past 2 months) a vector (4.4kb)and an insert(4.6kb) - both digested with XhoI enzyme. The colonies i get after transformation have all been negative. All sorts of troubleshooting has been done- vector is dephosphorylated, increased the vector: insert ration, enzymes and competent cells are functional, the insert and vector was confirmed with AGE, tried changing temperature of ligation. Still the ligation has failed. I have no clue what to do now. Please help ㅠㅠ