Hi Neha

I think your easiest option is to do blunt-end cloning. Double-digest your fragment with XhoI and SacII, digest your vector with XhoI, then make all the ends blunt by using Klenow + dNTPs. After that, dephosphorylate your cut vector with CIP or SAP to prevent it self-ligating. Just be careful to screen your clones for the correct orientation of insertion.

Alternatively you could design PCR primers for your insert with XhoI sites on both ends. Then it's just a single digest with no Klenow step (but still dephosphorylate your cut vector).

Good luck!

Hi Neha

I think your easiest option is to do blunt-end cloning. Double-digest your fragment with XhoI and SacII, digest your vector with XhoI, then make all the ends blunt by using Klenow + dNTPs. After that, dephosphorylate your cut vector with CIP or SAP to prevent it self-ligating. Just be careful to screen your clones for the correct orientation of insertion.

Alternatively you could design PCR primers for your insert with XhoI sites on both ends. Then it's just a single digest with no Klenow step (but still dephosphorylate your cut vector).

Good luck!

Thanks a lot Michael! I will try doing and let you know

I would agree with what Micheal said. I would add a little. 

You could also try to introduce the cuttinf site into the plasmid with PCR mutagenesis if its not too long. 

But with blunt end ligation you will not have control on direction of integration of your fragment. If you want the insertion to be direction specific then introduce the relative cutting site on either insert or Plasmid.