ligation

[Basil_S.Shorrosh]

   Hi,

   I have been trying to ligate:
       1-  A 7.5 kb SmaI fragment into a 7.5 SmaI digested vector.  Digestion
           was done at room temperature.
       2-  A 7.5 kb  ASP718/BamHI fragment into a 10 kb baculovirus vector
           and a 13 kb plant binary vector.

   Vectors were dephosphorylated.  Before ligation, inserts and and vectors
   were purified after running in an agarose gel using GeneClean kit.

   I have tried several conditions but with no success.

   If anyone have experience in cloning large fragments into large vectors, I
   would appreciate any suggestions.

   Thaks,  Basil